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Biomedical subjects

C Lindqvist

Publications and source records attributed to C Lindqvist.

At least 19 recordsLinked to original sources

Generation of a new protein purification matrix by loading ceramic hydroxyapatite with metal ions--demonstration with poly-histidine tagged green fluorescent protein.

The gene encoding the green fluorescent protein (GFP) from the jellyfish Aequorea victoria, was inserted under transcriptional control of the polyhedrin promoter of the Autographa californica nuclear polyhedrosis virus and expressed in the Spodoptera frugiperda insect cell line Sf9 during viral infection. The baculovirus transfervector pBlueBacHisB was used for constructing the recombinant baculovirus, so that the green fluorescent protein could be tagged with a poly-histidine tail. This fusion protein was utilized as a marker for evaluating the properties of metal ion loaded ceramic hydroxyapatite as a matrix in protein purification. Ceramic hydroxyapatite loaded with Zn(II) was the best choice for purifying this poly-histidine tagged GFP, followed by Fe(III) of the metal ions tested. Ni(II) that is superior especially in many poly-histidine purification systems did not, when loaded to hydroxyapatite, have binding properties comparable to Zn(II) or Fe(III). Elution of poly-histidine tagged GFP was best performed with phosphate buffers or EDTA that could compete with the phosphate molecules in hydroxyapatite or complexly bind the metal ions, respectively.

Animals

A microplate-based fluorometric assay for monitoring human cancer cell attachment to cortical bone.

A microplate-based fluorometric assay was developed for quantitation of cancer cell attachment to bone matrix. To evaluate this model system we used a panel of human cancer cell lines, including the human breast cancer cell line MDA-MB-231. For the assay, bovine cortical bone from the shaft of the femur was cut, turned, and sliced to 6-mm-diameter round 200-microm-thin disks and placed into the wells of a 96-well microplate. A fluorescent dye 2', 7'-bis(2-carboxyethyl-5(6)-carboxyfluorescein (BCECF-AM) and a microplate fluorometer equipped with a standard filter set for fluorescein isothiocyanate were used to detect the cells attached to the bone disks. The fluorescence was enhanced during the measurement step by using a K+ solution (pH 8.0) containing the H+/K+ ionophore nigericin, which equilibrates internal and external pH. By taking advantage of the pH sensitivity of BCECF, the fluorescence intensity in the assay was increased 2.5 times compared to cells loaded with calcein. The specificity of the assay was demonstrated with a specific immuneserum raised against the MDA-MB-231 cell line. The assay can be completed in 1 h and permits the use of a large number of samples and is therefore useful for screening potential drug candidates that could block cancer cell attachment to bone material. Moreover, the assay described can easily be used to characterize molecular structures involved in cell attachment to bone.

Animals

Histological study of tissue reactions to epsilon-caprolactone-lactide copolymer in paste form.

In previous studies, epsilon-caprolactone-lactide copolymer in solid form has been used in experimental animals as suture material, and as a biodegradable nerve guide. The aim of the study reported here was to assess tissue reactions to epsilon-caprolactone-lactide copolymer in paste form, histologically, and to compare bone healing at the sites of implantation versus that at control sites. The other purpose of the study was to evaluate the properties of the implanted material as a filling material for bone defects. Resorption time and intensity of inflammatory reaction were also evaluated. Material was implanted into the abdominal walls and femurs of 34 rats. Follow-up times were from 2 weeks to 1 year. The results showed that epsilon-caprolactone-lactide copolymer in paste form induces a severe inflammatory reaction when placed in muscle, and moderate inflammation when implanted into bone. The resorption time was more than 1 year. Bone healing at sites of implantation was slower than at control sites.

Animals

Lag-screw fixation of anterior mandibular fractures using biodegradable polylactide screws: a preliminary report.

PURPOSE: The purpose of this study was to evaluate the utility of biodegradable, self-reinforced poly-L-lactide screws (SR-PLLA) for lag-screw fixation of anterior mandibular fractures. PATIENTS AND METHODS: SR-PLLA lag-screws were used to stabilize anterior mandibular fractures in 11 patients. Maxillomandibular fixation was used to treat concomitant mandibular condyle fractures for 2 weeks in four patients and for 1 and 5 weeks in two patients. Clinical and radiologic follow-up lasted for 6 months in 36% of patients and for 1 year in 64%. RESULTS: Healing of all anterior fractures was uneventful, with no displacement or delay of bony union. No adverse reactions to the biodegradable screws were seen during follow-up. CONCLUSION: Biodegradable SR-PLLA screw fixation seems to be a new and promising way of treating anterior mandibular fractures.

Absorption

Fixation of mandibular body osteotomies using biodegradable amorphous self-reinforced (70L:30DL) polylactide or metal lag screws: an experimental study in sheep.

Mandibular body osteotomies were fixed in nine sheep using new totally amorphous (70L:30DL), self-reinforced, polylactide (SR-PLA) lag screws and in nine sheep using standard stainless steel lag screws. No intermaxillary fixation was used. During follow-up, radiological, histological and microradiological studies were undertaken at 3, 6, 12 and 24 weeks. In both groups, all osteotomies consolidated at similar rates and no adverse reaction to the screws was seen. However, displacements of the fixed osteotomy fragments were common in both groups during the first 3 weeks. The biocompatibility of SR-PLA during the follow-up period was found to be good. Only initial signs of biodegradation were seen. The results of this study indicate that (70L:30DL) SR-PLA has potential for use as a fixation screw material in oral and maxillofacial surgery, and that further studies using this material are justified.

Absorbable Implants

Osteotomy site healing following mandibular sagittal split osteotomy and rigid fixation with polylactide biodegradable screws.

Follow-up of 47 patients, treated using mandibular bilateral sagittal split osteotomy and self-reinforced poly-L-lactide acid (SR-PLLA) screws for rigid internal fixation, is presented. The focus was on clinical and radiological osteotomy healing. The average follow-up time was 2.1 years (range 0.5-5 years). Clinical recovery and radiological osteotomy healing during follow-up were uneventful. Osteolytic changes were seen around the SR-PLLA screws in 27% of cases. The majority of the screw canals remained as radiolucent shadows without bony filling.

Absorbable Implants

The role of alpha2 beta1 and alpha3 beta1 integrin receptors in the initial anchoring of MDA-MB-231 human breast cancer cells to cortical bone matrix.

Molecular mechanisms involved in the rapid attachment of the human breast cancer cell line MDA-MD-231 to cortical bone matrix were studied. The attachment of MDA-MD-231 cells to cortical bone disks could be blocked by 75% when cells were pretreated with a monoclonal antibody to the beta1-subunit of the integrin family. Monoclonal antibodies against the alpha2, alpha3, and alpha5 integrin subunits inhibited the attachment by 76, 26, and 8 % respectively. Collagenase type I and collagen type I antibody blocked the cell attachment by 45 and 50 % whereas pretreatment of the cells with soluble collagen type I blocked the attachment by 85 %. Our study with a panel of cancer cell lines further showed a close correlation between alpha2 beta1 and alpha3 beta1 integrin receptor expression and the capability to attach to cortical bone. These receptors appear to be the key receptors utilized by cancer cells for the initial attachment to cortical bone, and this could facilitate the localization of alpha2 beta1 and alpha3 beta1 expressing cancer cells to the skeleton.

Animals

Homogeneous time-resolved IL-2-IL-2R alpha assay using fluorescence resonance energy transfer.

A homogeneous receptor-ligand assay based on fluorescence resonance energy transfer is described. In the assay, recombinant human interleukin 2 (IL-2) and a monoclonal antibody against the human IL-2 receptor alpha chain were labelled with a highly fluorescent europium chelate and Cy5, respectively. As a result of a successful receptor-ligand complex formation, these labels are brought into close proximity, which will thereby allow an energy transfer to occur from the donor (europium) to the acceptor (Cy5), upon excitation of the donor. Utilization of specific non-neutralizing antibodies made it possible to use crude hIL-2R alpha membranes prepared from recombinant baculovirus-infected Sf9 insect cells. The specific energy transfer was measured at the emission wavelength of Cy5 using a time-resolved fluorometer. The data presented, demonstrate that this assay design can be utilized for saturation as well as competitive binding experiments in addition to regular receptor titrations. As a rapid simple homogeneous assay it is particularily suitable for high throughput screening analyses.

Animals

A 5-year in vitro and in vivo study of the biodegradation of polylactide plates.

PURPOSE: The purpose of this study was to investigate the long-term tissue response and duration of degradation of self-reinforced poly-L-lactide (SR-PLLA) multilayer plates in vivo. MATERIALS AND METHODS: Mandibular osteotomies in sheep were fixed with SR-PLLA multilayer plates. The animals were followed for 1, 2, 3, 4, and 5 years, after which histologic studies were performed. RESULTS: The foreign-body reaction was mainly mild, and the osteotomies were well united. After 5 years in vivo, the material was almost completely resorbed, but small particles of polymer could still be detected at the implantation site. SR-PLLA plates were also incubated in vitro for 5 years. The material degraded considerably faster in vivo than in vitro. Molecular weight, melting temperature, and crystallinity of the plates remained at a constant level after 2 years in vitro, indicating very slow degradation of the oligomeric (molecular weight [Mw], 3500 daltons), highly crystalline (heat of fusion, 70 J/g), PLLA residue solely as a result of hydrolysis. Although the plates became increasingly fragile as they degraded, they retained their macroscopic form until the end of the 5-year follow-up. Loss of mass of the plates was 52%+/-8% after 5 years of incubation in vitro. CONCLUSIONS: Although the long degradation period may seem to be a minor drawback to the use of such plates, it does not appear to affect the healing process.

Absorption

Occlusal and temporomandibular joint disorders in patients with unilateral condylar fracture. A prospective one-year study.

Adult dentate patients with unilateral condylar fractures were followed up for one year after injury. Patients were interviewed about subjective complaints, and mandibular excursions and function of occlusion and temporomandibular joints (TMJs) were recorded. Radiological characteristics were assessed from panoramic and Towne's views and compared between patients with occlusal disorders and patients without. During follow up, mandibular deviation on opening towards the fractured side was more pronounced in cases with marked reduction of ramus height and condylar dislocation. This restriction of translation movement of the fractured joint was also seen radiologically in one-third of cases while in two-thirds of the fractured condyles, malpositioning was observed when compared with the healthy side. In patients (39%) having subjective symptoms such as TMJ pain, altered occlusion or ability to bite only unilaterally, and objectively verified occlusal interferences, a marked reduction of the ramus height on the fractured side was observed. In such cases nonoperative treatment of condylar fractures may be compromised.

Adolescent

Skeletal stability following mandibular advancement and rigid fixation with polylactide biodegradable screws.

Skeletal stability during the first year after mandibular advancement surgery and fixation using biodegradable self-reinforced poly-l-lactide (SR-PLLA) screws, without postoperative intermaxillary fixation, was studied in 25 patients by means of cephalometric measurements. The magnitude of advancement was on average, 3.88 mm at pogonion (PG) (range 1.25-6.5 mm) and 4.57 mm at B-point (range 2.75-7.5 mm). After one year a mean relapse backwards of 0.59 mm at the PG (15%) and 0.78 mm at the B-point (17%) was observed. Nineteen patients (76%) and 17 patients (68%) were stable at PG and B-point, respectively. SR-PLLA screws are considered to be comparable to other forms of rigid internal fixation for fixation of bilateral splitting osteotomies after mandibular advancement, as far as skeletal stability is concerned.

Absorption

Biodegradable polylactide plates and screws in orthognathic surgery: technical note.

In orthognathic surgery, the bone fragments are usually fixed with metallic plates and screws. Metallic devices other than titanium plates are usually removed after the osteotomy has consolidated, which often requires general anaesthesia. Titanium plates, supposed to be biotolerable, have been introduced in order to overcome this need for secondary intervention. However, due to corrosion, titanium particles have been found in scar tissue covering these plates and in locoregional lymph nodes. Therefore, their removal is also advocated. Self-reinforced poly (L-lactide) homopolymer (PLLA) and poly (L/D-lactide) stereocopolymers with a L/D molar ratio up to 85/15 have sufficient strength to overcome the need for additional support for the fixation of fractures. The plates can be bent at room temperature. The surgical technique and early results of a case of bimaxillary surgery and genioplasty fixed with bioresorbable material without postoperative rigid maxillomandibular fixation are reported.

Absorption

Radiologic changes of the temporomandibular joint after condylar fractures in childhood.

OBJECTIVE: The purpose of this investigation was to study the radiologic outcomes of pediatric condylar fractures. STUDY DESIGN: Radiographs of 37 patients with 45 condylar fractures that had been diagnosed at the Department of Oral and Maxillofacial Surgery, Helsinki University Central Hospital were retrospectively reviewed. The frequency and characteristics of incomplete remodeling at an average of 4.1 years after the injury were studied. The influence of age and fracture type on remodeling were considered. RESULTS: Incomplete remodeling was observed after 56% of the fractures. The most frequent signs of incomplete remodeling were alteration in the configuration of the surface of the condylar head and deformation of the condylar neck. Incomplete remodeling was frequently (83%) related to fracture dislocation. For this fracture type the radiologic prognosis seems poor in all age groups. A difference in ramus height between the 2 sides was observed in 52% of the fractures, particularly frequently (80%) after fracture dislocation. CONCLUSIONS: Radiologic aberrations are frequent after pediatric condylar fractures. Dislocated fractures, in particular, need special attention and long-lasting clinical follow-up.

Adolescent

Malpractice claims for permanent nerve injuries related to third molar removals.

On the basis of the register of the Finnish Patient Insurance Association, the aim of this study was to examine malpractice claims for nerve injuries associated with third molar removals and determine whether they are concentrated among specialists, among less experienced dentists, or in certain geographic areas. During 1987-93 there were 139 claims for permanent sensory or motor disturbances related to removal of lower third molars in Finland. The lingual nerve was injured in 54% and the inferior alveolar nerve in 41% of the claims. In 91% of the cases the injury occurred in relation to surgical removal of the tooth and in 6% in relation to simple extraction. The claims were distributed among 123 dentists, of whom 78% were dental surgeons, 15% specialists in oral and maxillofacial surgery, and 7% other specialists. These figures represented 2% of the dental surgeons and 26% of the oral surgeons in Finland (P< 0.01). More than half the claims were associated with dentists with less than 10 years' experience. Claims originated more often from the eastern and northern (rural) areas of Finland than from urban areas (3.8 claims versus 2.4 claims per 100,000 inhabitants, P < 0.05). Compensation was paid to the patients in two-thirds of the cases, indicating that the dentists authorized to decide claims very often considered these injuries avoidable. Therefore, proper diagnosis, treatment planning, surgical techniques, and detailed patient information must be emphasized. In cases where risks are obvious, referral to an oral surgeon is recommended.

Adolescent

Inhibition of matrix metalloproteinase-1 by dichloromethylene bisphosphonate (clodronate).

Interstitial collagenase present in human jaw cyst extract and purified human fibroblast-type collagenase (MMP-1) were both efficiently inhibited in vitro by clodronate, an osteoactive, antiresorptive bisphosphonate. The IC50 of clodronate to inhibit MMP-1 is 150 microM. These findings suggest an extended and hitherto undescribed properties for clodronate/biphosphonates in prevention and treatment of tissue degradation in both bone and soft tissue destructive diseases.

Bone Diseases

Solid phase IL-2-IL-2R alpha assay with time resolved fluorometry.

A time-resolved fluorometric, solid phase, receptor ligand interaction assay is described. The assay consists of wells coated with anti-human IL-2 receptor alpha (hIL-2R alpha) monoclonal antibodies (mAb), europium labelled hIL2 (Eu-IL-2) and human recombinant IL-2 receptor alpha subunits expressed in the baculovirus expression vector system (BEVS). In the assay hIL-2R alpha-Eu-IL-2 complexes bind to the solid phase mAb. Receptor bound Eu is dissociated into an enhancer solution where it forms highly fluorescent complexes. The fluorescence is measured in a time-resolved fluorometer. The Kd value calculated from the saturation curve is in good agreement with previously reported values for the low affinity type of IL-2R, making the described assay a simple and nonradioactive alternative for measurement of soluble hIL-2R alpha in biological systems. Furthermore this assay format provides convenient separation of bound ligand from unbound and is therefore suitable for high throughput screenings.

Europium

An epidemiological study of patterns of condylar fractures in children.

OBJECTIVE: To find out how the site and type of condylar fracture are affected by its aetiology and the age and sex of the patient. DESIGN: Retrospective study. SETTING: Tertiary referral centre, Finland. MATERIAL: Radiographs of 101 children (aged 15 years or less) with 119 condylar fractures. MAIN OUTCOME MEASURES: Site of fracture and degree of displacement. RESULTS: A total of 26 of the 119 fractures were intracapsular (22%) and 93 (78%) extracapsular. Only among patients less than 6 years of age was there a preponderance of intracapsular fractures (7/12 fractures in 10 patients, 58%). In the older children 78% (83/107) were in the condylar neck. There were few subcondylar fractures (5/119, 4%). Only 6 fractures were displaced (5%). Dislocation of the condyle from the glenoid fossa was common in all age groups. CONCLUSION: The site of condylar fracture is age related, but not associated with sex or aetiology.

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