Biomedical subjects

Gary Balian

Publications and source records attributed to Gary Balian.

24 records · Page 2Linked to original sources

PubMed2003

Repair of full-thickness cartilage defects in rabbit knees with free periosteal graft preincubated with transforming growth factor.

This study compared different concentrations of transforming growth factor-beta 1 (TGF-beta1) used for short-term preincubation in vitro of large periosteal explants to determine the effect of chondrogenesis and the fate of repair cartilage over time. Eighty-two rabbits were divided into four groups: group A, non-grafted; group B, non-incubated grafted; group C, 100 ng/mL recombinant human (rh) TGF-beta1 preincubated grafted; and group D, 20 ng/mL rhTGF-beta1 preincubated grafted. Rabbits from each group were sacrificed at intervals between 2 and 24 weeks. Histologic slides were stained with safranin O and were scored based on a subjective scoring system. Group A healed with non-cartilaginous material. Group B healed with hyaline cartilage-like material with progressive thinning of this regenerated layer; at 24 weeks, this layer was fibrous tissue. Group C enhanced repair with hyaline cartilage-like material but accelerated early degeneration and osteophyte formation; the cartilage became fibrous tissue at 24 weeks. Group D did not enhance cartilaginous repair. At 24 weeks, all groups had the same result. The 100 ng/mL rhTGF-beta1 preincubation in vitro with periosteum enhanced early osteochondral repair but did not show prolonged durability. Clinical application of this growth factor necessitates further study.

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PubMed2002

Transplanted bone marrow cells localize to fracture callus in a mouse model.

Bone marrow contains many cellular elements that may contribute to fracture repair. We used a pluripotential stromal cell in a mouse model to demonstrate the presence of transplanted cells in fracture hematoma and subsequently in maturing fracture callus. Cells were transduced with traceable genes (lac Z and neomycin resistance) and traced in vivo after intravenous injection into syngeneic mice. These transduced cells home to bone marrow, suggesting that they might be detected in fracture callus. Cells were injected intravenously into mice and stabilized femoral shaft fractures were induced. Control mice received intravenous lactated-Ringer's solution prior to fracture. Callus tissue and marrow were examined histologically from I to 10 weeks after fracture to detect transplanted cells. Transplanted cells were detected in fracture callus in areas, and at times, of most active bone formation. Control specimens showed minimal staining of the callus tissue. Levels of the traceable gene in fracture callus increased, reached a peak between 3 and 4 weeks after fracture, then diminished and disappeared by 10 weeks post-fracture as woven bone at the fracture site was replaced by lamellar bone with cells from the host mouse. The results show that pluripotent bone marrow cells home to the marrow after systemic injection and localize in fracture callus.

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PubMed2002

Transforming growth factor-beta in calcium alginate beads for the treatment of articular cartilage defects in the rabbit.

PURPOSE: Articular cartilage has only limited capability for intrinsic repair. The use of growth factors has been suggested to improve the repair of cartilage after injury. Reliable delivery systems for these agents are needed. In this study we tested calcium alginate for the delivery of TGF-beta in the treatment of osteochondral defects in the rabbit knee. TYPE OF STUDY: Randomized trial animal study and basic science study. METHODS: In vitro, to establish the kinetics of TGF-beta release from the alginate, 125I- labeled TGF-beta was suspended in 1.2% sodium alginate at concentrations of 1 microg/mL and 10 microg/mL. Beads were formed from 50 microL aliquots and placed into standard culture medium by immersion in calcium chloride solution and incubated at 37 degrees C. A gamma counter was used to measure the amount of TGF-beta that was released into the medium at various time points. In vivo, osteochondral defects were created in the trochlear grooves of 32 New Zealand White rabbits. Defects were treated with plain alginate or with alginate containing TGF-beta at 20 ng/mL or 2,000 ng/mL. Untreated defects served as a control. Animals were killed after 6 and 12 weeks. Knee joints were evaluated grossly with a 12-point grading scale. Histologic sections of the repair tissue were stained with Safranin O and evaluated using a 24-point grading scale by 2 independent blinded observers. Mean scores and standard deviations were calculated. P values were determined using the Student t test. RESULTS: The TGF-beta was released at a surprisingly slow but steady rate. Release rates extrapolated from the gamma counter measurements were 0.25% per hour and 0.33% per hour, for the 1 microg/mL and 10 microg/mL beads, respectively. Gross analysis scores at 6 and 12 weeks resulted in higher scores for both TGF-beta groups without reaching statistical significance. The lower TGF-beta concentration reached the highest scores, whereas the higher concentration (2,000 ng/mL) resulted in increased osteophyte formation. Histologic analysis at 6 weeks resulted in average scores ranging from 14.5 for empty defects and 18.1 for alginate-treated defects, to 20.0 and 20.3 for the 2,000 ng/mL and 20 ng/mL TGF-beta groups, respectively (P <.05). At 12 weeks, histologic scores ranged from 14.9 for empty and 14.5 for alginate to 20.1 and 20.5 for the 2,000 ng/mL and 20 ng/mL TGF-beta groups, respectively (P <.05). These results indicate a significant improvement of the quality of the repair tissue at 6 and 12 weeks with TFG-beta treatment, especially at the lower concentration. CONCLUSIONS: The use of alginate allows the controlled delivery of TGF-beta selectively to the site of injury, potentially avoiding systemic side effects. Furthermore, treatment with TGF-beta appears to improve the repair of articular cartilage defects. Longer-term studies are needed to assess whether the benefits of the TGF-beta treatment can be sustained.

Alginates↗

PubMed2002

Systemically administered mesenchymal stromal cells transduced with insulin-like growth factor-I localize to a fracture site and potentiate healing.

OBJECTIVE: To determine to the ability of systemically administered pluripotential mesenchymal stromal cells to localize to a fracture site and whether transduction with a therapeutic gene, insulin-like growth factor-I (D1-IGF-I), could potentiate healing. DESIGN: Murine model, basic science study. SETTING: Laboratory. SPECIMENS: Closed, transverse, mid-shaft femur fractures were produced in 108 Balb/c mice after intramedullary stabilization. INTERVENTIONS: A cloned, pluripotential, mesenchymal cell line, termed D1, was stably transfected with either the gene beta-galactosidase (D1-BAG) as a histologic marker or with the gene IGF-I (D1-IGF-I) growth factor. Mice received systemic injections of either D1-BAG cells for in vivo localization or D1-IGF-I for therapeutic intervention. A third group received lactated Ringer's solution and served as control. MAIN OUTCOME MEASUREMENTS: Sections obtained from the fracture site and contralateral femurs were examined histologically and by deoxyribonucleic acid-polymerase chain reaction (DNA-PCR) to detect the presence of transplanted cells at 2, 4, and 6 weeks after fracture. Matrix mineralization and callus maturation were evaluated by histology. RESULTS: At all time points, using histologic staining with X-gal and deoxyribonucleic acid-polymerase chain reaction for marker genes, there was a statistically greater number of transplanted cells ( p< 0.001) and significantly higher DNA-PCR for marker genes ( p< 0.001) in the fractured femurs than in the nonfractured femurs. Mice receiving D1-IGF-I cells demonstrated a greater percent of mineralized callus than controls at two weeks (p < 0.05). At 4 and 6 weeks, treated mice demonstrated on average greater mineralized matrix and accelerated progression to an osseous callus as compared with the control group. CONCLUSIONS: Cell-based gene therapy has the potential to deliver higher therapeutic levels of growth factors specifically at the site of cell localization while minimizing wider systemic side effects. This study demonstrates that systemically injected IGF-I transduced, mesenchymal cells are able to return to and repopulate the bone marrow. More importantly, these cells localize preferentially to a fracture site and accelerated fracture healing.

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PubMedDate not supplied

Tendon tissue engineering and gene transfer: the future of surgical treatment.

Technologic improvements in the field of tissue engineering are leading to new potential developments in the currently used approaches to treat tendon injuries including difficult clinical scenarios such as zone II flexor tendon injuries of the hand and the mutilated hand with extensive tendon defects. A combination of mesenchymal (adult stem) cells, growth factors, and bioresorbable polymers can provide a solution for the treatment of difficult tendon injuries. Extensive research is needed to show that the extracellular matrix produced in response to the cell/growth factor/polymer composites in vivo is effective and functional as a regenerate tissue. Further exciting advances are foreseen in cell-based genetic engineering with the transfer of DNA to the site of tendon lacerations. These treatment modalities require improved safety precautions to reduce the risks and enhance the benefits of gene therapy.

Animals↗

PubMedDate not supplied

Tracing transduced cells in osteochondral defects.

In this study the authors explored the feasibility of using transduced cells for gene therapy to induce healing of osteochondral defects. Both a mouse mesenchymal cell line and mixed rabbit adherent stromal cells were transduced with either liposomal transfection or retroviral transduction using a traceable gene. Transduction efficiency was more than 95% with the retroviral construct and expression was maintained for over 6 months of passage. The liposomal transfection led to a transient expression with an efficiency of 50%. The expression of osteochondral genes was diminished but preserved after transduction in vitro. Transduced rabbit cells were transplanted into osteochondral defects in rabbit femoral condyles. Cells transplanted in vivo could be detected for 4 weeks in the repair tissue. The authors' data demonstrate that mesenchymal cells from bone marrow, stably transduced with a traceable gene product, retain the bone and cartilage phenotype and can be followed in vivo after transplantation into cartilage defects.

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