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Biomedical subjects

Brian J F Wong

Publications and source records attributed to Brian J F Wong.

At least 19 recordsLinked to original sources

Stress relaxation in porcine septal cartilage during electromechanical reshaping: mechanical and electrical responses.

Electromechanical reshaping (EMR) of facial cartilage has recently been developed as an alternative to classic surgical techniques to alter cartilage shape. This study focuses on determining the underlying physical mechanisms responsible for shape change (stress relaxation) in mechanically deformed facial cartilage specimens exposed to constant electric fields. Flat porcine nasal septal cartilage specimens were deformed by an aluminum jig into semicylindrical shapes while a constant electric voltage was applied to the concave and convex surfaces of the specimen. Mechanical stress, electric current and resistance were measured during voltage application. Specimen shape retention was measured as retained bend angle. Total electric charge transferred in the electric circuit was calculated from the electric current measurement. Electrical resistance, transferred charge and the bend angle increase with increase in voltage application time until bend angle reaches maximum value determined by the jig geometry. Then, the bend angle decreases and electrical parameters nearly saturate. The time dependent behavior of electric current was analyzed using the Cottrell equation. The observed changes in electric current suggest that during the initial 1-2 min of EMR nonlinear diffusion determines electro-chemical reaction rates, which are then followed by a linear diffusion dominated process. Close correlation between alteration of cartilage mechanical state and change in its electrical properties suggest that an electro-chemical reaction is the dominant mechanism behind EMR.

Animals↗

Noninvasive measurement of ablation crater size and thermal injury after CO2 laser in the vocal cord with optical coherence tomography.

OBJECTIVE: To characterize tissue destruction after CO(2) laser-ablation of the vocal cords with the use of optical coherence tomography (OCT). STUDY DESIGN AND SETTING: OCT was used to image fresh porcine vocal cords after laser ablation. OCT and histology estimates of the ablation crater dimensions and the depth of thermal injury were obtained. RESULTS: The vocal cord substructures up to 2.29 mm in depth at 10 microm resolution, and the thermal disruption after laser ablation were identified by OCT. OCT and histology estimates of the lesion dimensions showed no significant differences. Crater depth is directly proportional to laser power, whereas crater width and the zone of thermal injury appear to be unrelated to laser power. CONCLUSIONS: OCT may be used to accurately characterize the native states and the laser-induced thermal injury of laryngeal mucosa, within the inherent limitation in its depth of penetration. OCT may be a useful diagnostic and monitoring tool in an otolaryngology practice.

Animals↗

Optical coherence tomography of laryngeal cancer.

OBJECTIVES: Optical coherence tomography (OCT) is a high-resolution optical imaging technique that produces cross-sectional images of living tissues using light in a manner similar to ultrasound. This prospective study evaluated the ability of OCT to identify the characteristics of laryngeal cancer and measure changes in the basement membrane, tissue microstructure, and the transition zone at the edge of tumors. MATERIALS AND METHODS: One hundred thirty-three patients underwent OCT examination during surgical endoscopy of the head and neck. Twenty-two patients with laryngeal cancer or a history of laryngeal cancer were imaged with a fiberoptic OCT system. Tumor and adjacent transition zones were imaged along with uninvolved subsites. OCT images were correlated with histopathology. RESULTS: Twenty-six OCT examinations were performed in 22 patients. Basement membrane disruption was seen in 18 subjects, all of whom had histology showing classic features of cancer. A transition zone to uninvolved epithelium at the tumor periphery was also often observed. In six studies, benign or premalignant processes were histologically confirmed. In three thin, superficial lesions, an intact basement membrane was observed. The basement membrane could not be identified in three other bulky exophytic, premalignant lesions, primarily because of increased superficial signal backscattering observed in pathologic tissues. CONCLUSIONS: OCT clearly identifies basement membrane violation from laryngeal cancer and can identify transition zones at the cancer margin. In bulky exophytic lesions, OCT signal may not penetrate deeply enough to show the basement membrane, but for many suspicious lesions that require exclusion of cancer, OCT shows potential for assisting in diagnostic assessment.

Aged↗

Depth-resolved phase retardation measurements for laser-assisted non-ablative cartilage reshaping.

Since polarization-sensitive optical coherence tomography (PS-OCT) is emerging as a new technique for determining phase retardation in biological materials, we measured phase retardation changes in cartilage during local laser heating for application to laser-assisted cartilage reshaping. Thermally-induced changes in phase retardation of nasal septal cartilage following Nd:YAG laser irradiation were investigated using a PS-OCT system. A PS-OCT system and infrared imaging radiometer were used to record, respectively, depth-resolved images of the Stokes parameters of light backscattered from ex vivo porcine nasal septal cartilage and radiometric temperature changes following laser irradiation. PS-OCT images of cartilage were recorded before (control), during and after laser irradiation. From the measured Stokes parameters (I, Q, U and V), an estimate of the relative phase retardation between two orthogonal polarizations was computed to determine birefringence in cartilage. Phase retardation images of light backscattered from cartilage show significant changes in retardation following laser irradiation. To investigate the origin of retardation changes in response to local heat generation, we differentiated two possible mechanisms: dehydration and thermal denaturation. PS-OCT images of cartilage were recorded after dehydration in glycerol and thermal denaturation in heated physiological saline. In our experiments, observed retardation changes in cartilage are primarily due to dehydration. Since dehydration is a principal source for retardation changes in cartilage over the range of heating profiles investigated, our studies suggest that the use of PS-OCT as a feedback control methodology for non-ablative cartilage reshaping requires further investigation.

Animals↗

Low-voltage polymer-based scanning cantilever for in vivo optical coherence tomography.

Novel hand-held optical coherence tomography (OCT) probes with polymer cantilevers have been developed for clinical oral and skin imaging. An electroactive ionic polymer-metal composite cantilever was used to generate 3-mm transverse scanning movement of an optical fiber with applied 2-V linear alternating voltage at 1 Hz. Low driving voltage ensures safety. Two different optical designs achieve both forward and sidewise scanning and make it possible to image everywhere within the human oral cavity. In vivo OCT imaging of the human tongue is demonstrated.

Equipment Design↗

Characterization of submucosal lesions using optical coherence tomography in the rabbit subglottis.

OBJECTIVE: To evaluate the efficacy of optical coherence tomography in differentiating between several simulated subglottic lesions, using an ex vivo rabbit laryngotracheal model. DESIGN: Laryngotracheal complexes were harvested from euthanized rabbits and divided into the following 4 groups: (1) control, (2) submucosal collagen injection (simulating scar formation), (3) dehydration and rehydration (simulating edema), and (4) repeated intubation trauma. The subglottic region was imaged using optical coherence tomography. Images were later correlated with conventional histologic findings. RESULTS: The epithelium, basement membrane, lamina propria, perichondrium, and cartilage (cricoid and tracheal) were clearly imaged. In group 2, an increase in the thickness of the lamina propria was observed, in addition to a characteristic optical pattern of the injected collagen. Dehydration (in group 3) produced a visible reduction in the thickness of the lamina propria, while rehydration of the same specimen with distilled water revealed a significant increase in submucosal swelling. Repeated intubation (in group 4) resulted in tissue edema that was seen as wavy heterogeneous thickening of the lamina propria. Edema produced by repeated intubation or distilled water immersion was easily differentiated from native and collagen-injected tissues. CONCLUSION: Optical coherence tomography successfully identifies the microstructure layers of the subglottis and can differentiate between edema and increased collagen deposition in the rabbit model.

Animals↗

Long-term in vivo stability of rabbit nasal septal cartilage following laser cartilage reshaping: a pilot investigation.

BACKGROUND AND OBJECTIVES: To evaluate the long-term effect of laser cartilage reshaping on rabbit nasal septal cartilage viability and mechanical integrity in an in vivo model. STUDY DESIGN/MATERIALS AND METHODS: In vivo animal investigation. Rabbit septal cartilage specimens were laser (Nd:YAG, lambda = 1.32 mum, spot size 5.4-mm diameter, 10 W, 10 seconds, 50 Hz PPR) reshaped and subsequently reimplanted into an interscapular subcutaneous pocket. Specimens were harvested at 8 and 12 months and evaluated using photography, flow cytometry, and histology. RESULTS: Grossly, specimens showed alteration in the physical integrity with varying degrees of tissue resorption. The non-irradiated control specimens demonstrated significantly increased stiffness. Histologically, there was marked depletion of the extracellular matrix and an overall reduction in tissue mass in laser irradiated tissues. However, flow cytometry data identified viable chondrocytes in laser-irradiated specimens that were identical to those observed in controls. CONCLUSIONS: Study results demonstrate that the rabbit nasal septal cartilage model can be effectively used to study laser reshaping, however alternative recipient sites with perichondrial lining, such as the pinna, may provide a more realistic physiologic environment for reshaped graft tissue. The dosimetry used in this pilot study likely led to significant thermal injury. Study results underscore the importance of elucidating the optimal laser dosimetry required to initiate permanent shape change while minimizing thermal damage.

Animals↗

Identification of chondrocyte proliferation following laser irradiation, thermal injury, and mechanical trauma.

BACKGROUND AND OBJECTIVE: Cartilage has a limited regenerative capacity, and there are a lack of reliable techniques and methods to stimulate growth of new tissue to treat degenerative diseases and trauma. This study focused on identifying chondrocyte cell proliferation in ex vivo cartilage tissue following heating Nd:YAG laser using whole-mount analysis and flow cytometry, and compared findings with results produced by contact, and water bath heating methods, mechanical injury, and the addition of transforming growth factor-beta (TGF-beta). STUDY DESIGN/MATERIALS AND METHODS: Ex vivo rabbit nasal septal cartilages were either irradiated with an Nd:YAG laser (lambda = 1.32 microm, 2-16 seconds, 6 W/cm(2)), heated by immersion in a warm saline bath, heated by direct contact with a metal rod, or mechanically damaged by scoring with a scalpel or crushing. After treatment, specimens were incubated for 7 or 14 days in growth media containing 10 microM bromodeoxyuridine (BrdU). Additional specimens were cultured with both BrdU and TGF-beta. Both whole-mount BrdU-double-antibody detection techniques and flow cytometry were used to determine the presence of DNA replication as a marker of proliferation. RESULT: An annular region of regenerating chondrocytes was identified surrounding the laser irradiation zone in whole-mount tissue specimens, and the diameter of this region increased with irradiation time. Using whole-mount analysis, no evidence of chondrocyte DNA replication was observed in tissues heated using non-laser methods, grown in TGF-beta, or mechanically traumatized. In contrast, flow cytometry identified the presence of BrdU-positive cells in the S-phase of the cell cycle (synthesis of DNA) for all protocols, indicating chondrocyte proliferation. The percentage of cells that are in S-phase increased with irradiation time. CONCLUSION: These data provide evidence that laser irradiation, along with other thermal and mechanical treatments, causes a proliferative response in chondrocytes, and this is observed ex vivo in the absence of cellular and humoral repair mechanisms. The advantage of using optical methods to generate heat in cartilage is that microspot injuries could be created in tissue and scanned across surfaces in clinical applications.

Animals↗

Photodynamic therapy on keloid fibroblasts in tissue-engineered keratinocyte-fibroblast co-culture.

BACKGROUND AND OBJECTIVES: Keloids are disfiguring, proliferative scars that are a pathologic response to cutaneous injury. An organotypic tissue culture system (the Raft model 1-10) was used to investigate the feasibility of using photodynamic therapy (PDT) as an adjunctive therapy to treat keloids following surgical excision. The Raft co-culture system mimics skin by layering keratinocytes on top of fibroblasts embedded in a collagen matrix. PDT uses drugs that produce singlet oxygen in situ when irradiated by light, and may lead to a number of effects in living tissues varying from the modulation of growth to apoptosis. PDT is already used to treat several benign and malignant diseases in organs such as the skin, retina, and esophagus. STUDY DESIGN/MATERIALS AND METHODS: Normal adult, neonatal, and keloid fibroblasts and keratinocytes were isolated from skin obtained from patients undergoing elective procedures and used to construct the Rafts. Mature Rafts (after 4 days) were incubated with 5-amino levulinic acid (5-ALA), a photosensitizer, for 3 hours and were laser-irradiated (635 nm) for total energy delivery of 5 J/cm2, 10 J/cm2, or 20 J/cm2. Rafts were examined 24 hours and 14 days later. Cell viability was determined using confocal imaging combined with live-dead fluorescent dyes. Multi-photon microscope (MPM) imaged collagen structure and density. As Rafts contract over time, surface area was measured using optical micrometry daily. RESULTS: At 10 and 20 J/cm2, near-total cell death was observed in all constructs, while at 5 J/cm2 cell viability was comparable to controls. Cell viability in keloid and neonatal Rafts was greater than that observed in normal adult Rafts. Treated Rafts contracted less over the 14-day period compared to controls. Contraction and collagen density were greatest in keloid and neonatal Rafts. CONCLUSIONS: A PDT dosimetry range was established, which reduces tissue contraction and collagen density while minimizing injury to fibroblasts.

Adult↗

A novel laryngoscope instrument stabilizer for operative microlaryngoscopy.

OBJECTIVE: To evaluate and optimize the design of a removable and inexpensive internal stabilization device to reduce the effect of intention tremor during laryngeal microsurgery. STUDY DESIGN AND SETTING: In this laboratory investigation, stabilizers were designed and constructed to allow a nonobstructing view of the surgical field, permit simple insertion and removal, and accommodate microsurgical instruments. Prototype stabilizers were tested by using a Dedo laryngoscope, a measurement grid, and video recording equipment, which recorded instrument tremor within the magnified operative field for later analysis. Physicians also rated instrument stability, mobility, visualization, and ease of use on a survey form. RESULTS: Instrument tremor was reduced approximately 90%, with little obstruction of view of the surgical field. Instrument range of motion was reduced but improved rapidly as the stabilizer bar was moved further from the tip of the laryngoscope. CONCLUSIONS: Use of a stabilization device in the laryngoscope lumen reduces instrument tremor and has the potential to improve surgical performance during laryngeal microsurgery. EBM RATING: B-3.

Equipment Design↗

Measurement of morphologic changes induced by trauma with the use of coherence tomography in porcine vocal cords.

BACKGROUND AND OBJECTIVE: To evaluate the ability of optical coherence tomography (OCT) (a noncontact, high-resolution imaging technique that generates detailed cross-sectional images of tissue structure) to image and document acute response to trauma in the larynx. MATERIALS AND METHODS: Porcine larynges were dissected to expose the vocal cords. Vocal cords were imaged with 1310 nanometer (nm) light source with a bandwidth of 75 nm at 1 frame/second (s). Vocal cord injuries were simulated by applying topical phenolic acid, injecting titanium dioxide into the submucosa, and carbon dioxide laser irradiation. RESULTS: The epithelium, basement membrane, and lamina propria were clearly delineated, and the effect of each intervention could be monitored as lesions progressed. The OCT image corresponded closely with histology. CONCLUSION: OCT is a powerful imaging tool with the potential for use in real time and has potential for multiple clinical applications in the larynx.

Administration, Topical↗

In vivo optical coherence tomography of the human larynx: normative and benign pathology in 82 patients.

OBJECTIVES: Optical coherence tomography (OCT) is an emerging imaging modality that combines low-coherence light with interferometry to produce cross-sectional images of tissue with resolution about 10 mum. Patients undergoing surgical head and neck endoscopy were examined using a fiberoptic OCT imaging probe to study and characterize microstructural anatomy and features of the larynx and benign laryngeal pathology in vivo. STUDY DESIGN: Prospective clinical trial. MATERIALS AND METHODS: OCT imaging of the larynx was performed in 82 of 115 patients who underwent surgical endoscopy for various head and neck pathologies. The OCT device employs a 1.3 microm broadband light source (FWHM, 80 nm). The frame rate is 1 Hz. Imaging was performed using a handheld probe placed in near contact with the target site. The maximum axial and lateral dimensions for the region of interest imaged were 2.5 mm x 6 mm, with resolutions of 10 microm. Simultaneously, conventional endoscopic images were obtained to provide anatomic correlation with OCT images and histology. Optical micrometry was performed to measure the epithelium thickness. RESULTS: Systematic OCT imaging of laryngeal structures and subsites provided information on the thickness of the epithelium, integrity of the basement membrane, and structure of the lamina propria. Microstructural features identified included glands, ducts, blood vessels, fluid collection/edema, and the transitions between pseudostratified columnar and stratified squamous epithelium. The mean epithelial thickness of laryngeal subsites was calculated: true vocal cord (129 microm), false vocal cords (124 microm), aryepiglottic fold (177 microm), subglottis (98 microm), and epiglottis (185 microm). True vocal cord pathology imaged included Reinke's edema, papillomatosis, polyps, mucous cysts, and granulation tissue. Subglottic imaging identified boundaries between epithelium, lamina propria, and cartilage. The OCT images compared favorably with conventional histopathology. CONCLUSION: OCT has the unique ability to image laryngeal tissue microstructure and can detail microanatomic changes in benign, premalignant, and malignant laryngeal pathologies. OCT holds the potential to guide surgical biopsies, direct therapy, and monitor disease, particularly when office-based systems are developed. This is a promising imaging modality to study the larynx.

Biopsy↗

Lasers in pediatric airway surgery: current and future clinical applications.

BACKGROUND AND OBJECTIVES: The smaller anatomy and limited access to instrumentation pose a challenge to the pediatric airway surgeon. The enhanced precision and ability to photocoagulate tissue while operating with the laser enhances the surgeon's ability to successfully treat unique pediatric conditions. STUDY DESIGN/MATERIALS AND METHODS: A comprehensive literature review was performed using the Medline (1995-2004) database. RESULTS: Due to its shallow tissue penetration and thermal effect, the carbon dioxide (CO2) laser is generally considered the laser of choice for pediatric airway applications. The potential for increased scarring and damage to underlying tissue caused by the greater penetration depth and thermal effect of the Nd:YAG and potassium-titanyl-phosphate (KTP) lasers preclude their widespread use in this population. CONCLUSIONS: In this review, we will discuss the clinical applications of lasers in pediatric airway surgery, the current state of the art, where the current technology is deficient, and the potential applications of novel laser systems.

Child↗

Rate process analysis of thermal damage in cartilage.

Cartilage laser thermoforming (CLT) is a new surgical procedure that allows in situ treatment of deformities in the head and neck with less morbidity than traditional approaches. While some animal and human studies have shown promising results, the clinical feasibility of CLT depends on preservation of chondrocyte viability, which has not been extensively studied. The present paper characterizes cellular damage due to heat in rabbit nasal cartilage. Damage was modelled as a first orderrate process for which two experimentally derived coefficients, A = 1.2 x 10(70) s(-1) and Ea = 4.5 x 10(5) J mole(-1), were determined by quantifying the decrease in concentration of healthy chondrocytes in tissue samples as a function of exposure time to constant-temperature water baths. After immersion, chondrocytes were enzymatically isolated from the matrix and stained with a two-component fluorescent dye. The dye binds nuclear DNA differentially depending upon chondrocyte viability. A flow cytometer was used to detect differential cell fluorescence to determine the percentage of live and dead cells in each sample. As a result, a damage kinetic model was obtained that can be used to predict the onset, extent and severity of cellular injury to thermal exposure.

Animals↗

Quantitative assessment of chondrocyte viability after laser mediated reshaping: a novel application of flow cytometry.

BACKGROUND AND OBJECTIVES: Lasers can be used to reshape cartilage by accelerating mechanical stress relaxation. In this study, fluorescent differential cell viability staining and flow cytometry were used to determine chondrocyte viability following laser heating. STUDY DESIGN/MATERIALS AND METHODS: Porcine septal cartilages were irradiated with an Nd:YAG laser (lambda = 1.32 microm, 25 W/cm(2)) while surface temperature, stress relaxation, and diffuse reflectance were recorded. Each slab received one, two, or three laser exposures (respective exposure times of 6.7, 7.2, 10 seconds). Irradiated samples were then divided into two groups analyzed immediately and at 5 days following laser exposure. Chondrocytes were isolated following serial enzymatic digestion, and stained using SYTO/DEAD Red (Molecular Probes, Eugene, OR). A flow cytometer was then used to detect differential cell fluorescence; size; granularity; and the number of live cells, dead cells, and post-irradiation debris in each treatment population. RESULTS: Nearly 60% of chondrocytes from reshaped cartilage samples isolated shortly after one irradiation, were viable while non-irradiated controls were 100% viable. Specimens irradiated two or three times demonstrated increasing amounts of cellular debris along with a reduction in chondrocyte viability: 31 and 16% after two and three exposures, respectively. In those samples maintained in culture medium and assayed 5 days after irradiation, viability was reduced by 28-88%, with the least amount of deterioration in untreated and singly irradiated samples. CONCLUSIONS: Functional fluorescent dyes combined with flow cytometric analysis successfully determines the effect of laser irradiation on the viability of reshaped cartilage.

Animals↗

Characterization of temperature dependent mechanical behavior of cartilage.

BACKGROUND AND OBJECTIVES: Few quantitative studies have investigated the temperature dependent viscoelastic properties of cartilage tissue. Cartilage softens and can be reshaped when heated using laser, RF, or contact heating sources. The objectives of this study were to: (1) measure temperature dependent flexural storage moduli and mechanical relaxation in cartilage, (2) determine the impact of tissue water content and orientation on these mechanical properties, and (3) use these measurements to estimate the activation energy associated with the mechanical relaxation process. STUDY DESIGN/MATERIALS AND METHODS: Porcine nasal septal cartilage specimens (30 x 10 x 2 mm) were deformed using a single cantilever arrangement in a dynamic thermomechanical analyzer. Stress relaxation measurements were made at discrete temperatures ranging from 25 to 70 degrees C in response to cyclic deformation (within the linear viscoelastic region). The time and temperature dependent behavior of cartilage was measured using frequency multiplexing techniques (10-64 Hz), and these results were used to estimate the activation energy for the phase change using the Williams-Landel-Ferry (WLF) equation and the Arrhenius kinetic equation. In addition, the effect of tissue orientation was examined with specimens oriented in both transverse and longitudinal directions at room temperature. RESULTS: The storage moduli of porcine cartilage decreased with increasing temperature, and a critical change in mechanical properties was observed between 58 and 60 degrees C with a reduction in the storage modulus by 85-90%. The shift of the stress relaxation behavior from viscoelastic solid to viscoelastic liquid was observed between 50 and 57 degrees C and likely corresponds to the transition temperature region in which structural changes in the tissue occur. The storage moduli for transverse and longitudinally oriented specimens were 19-22 and 14-16 MPa, respectively at ambient temperature. Reducing the water content (<10% mass loss) by allowing it to dry under ambient conditions resulted in reduction in the storage modulus by 31-36%. The activation energy associated with the mechanical relaxation of cartilage was 147 kJ/mole at 60 degrees C. This value was calculated by measuring stress-strain relationship under conditions where linear viscoelastic behavior was observed (0.09-0.15% of strain) within the transition temperature region (58-60 degrees C). CONCLUSIONS: The anisotropic mechanical behavior of cartilage was quantitatively analyzed in the transversely and longitudinally oriented specimens. Viscoelastic behavior appeared to be strongly dependent on the water content. Using empirically determined estimates of the transition zone temperature range accompanying stress relaxation, the activation energy for stress relaxation was calculated using time and temperature superposition theory and WLF equation. Further investigation of the molecular changes, which occur during laser irradiation, may assist in understanding the thermal and mechanical behavior of cartilage and how the reshaping process might to be optimized.

Animals↗

Measurement of the elastic modulus of rabbit nasal septal cartilage during Nd:YAG (lambda = 1.32 microm) laser irradiation.

BACKGROUND AND OBJECTIVES: The objective of this study was to quantitatively measure changes in the elastic moduli of rabbit nasal septal cartilage during laser heating. While the efficacy of laser cartilage reshaping has been established for use in nasal surgery, few studies have investigated the temperature-dependent viscoelastic behavior of cartilage. STUDY DESIGN/MATERIALS AND METHODS: Cyclic force versus displacement curves were generated during the Nd:YAG laser (lambda = 1.32 microm, 10 second exposure time, 21.22 W/cm2) irradiation of cartilage specimens secured in cantilevered geometry. Samples were irradiated three times with 30 second cooling intervals between each laser exposure. Measurements were recorded before, during, and after laser irradiation, and then following complete rehydration in normal saline (NS) for 1 hour at 25 degrees C. Elastic modulus was calculated assuming linear viscoelastic behavior. RESULTS: The elastic modulus in native tissue decreased during and after successive laser exposures from about 6 to 3.5 MPa. After rehydration, the modulus returned to near-baseline value. Surface temperature reached a maximum of 65 degrees C. CONCLUSIONS: The laser irradiation of cartilage using parameters similar to those used in reshaping does not produce significant irreversible changes in the mechanical properties of the tissue. Measurement of the elastic modulus is an effective means of characterizing alterations in cartilage mechanical behavior during and after laser heating.

Animals↗