Scientific workshop on the biology and pathology of acquired connective tissue diseases.
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Arthritis, Rheumatoid↗
Biomedical subjects
V M Goldberg
Publications and source records attributed to V M Goldberg.
S-100 protein immunostaining identifies cells expressing a chondrocytic phenotype during articular cartilage repair.
The healing of articular surface defects has been studied with conventional histology, which relies on the staining of the extracellular matrix to identify the phenotype of the cells present. A chondrospecific cellular marker would be useful. S-100 protein has been found in all chondroid tissues studied, and we evaluated its usefulness in the study of articular cartilage repair. Full-thickness rabbit femoral condylar defects were made, and the specimens were studied at serial time intervals. S-100 protein staining positively showed chondroid cells in the 7- and 14-day specimens, which were not identifiable by conventional techniques. At 30 and 60 days, an S-100 positive band of cells separated a deep safranin-O positive hypertrophic layer from a fibrocellular surface layer. At 120 days, the presence of S-100 protein identified cells with chondrogenic potential, and the lack of S-100 protein in other cells embedded in conventionally stained matrix suggested that these cells were no longer of a chondroid phenotype. The presence of S-100 protein-identified chondroid cells early in the repair process when the cells had not begun to synthesize conventionally stainable matrix and the lack of S-100 protein in cells late in the repair positively identified a phenotypic change earlier than conventional histology.
Animals↗
The effects of vascularity and cyclosporin A on the mechanical properties of canine fibular autografts.
We studied the biomechanical behavior of orthotopic canine autografts as influenced by vascularized supply and the administration of cyclosporin A at three months and six months post-surgery. The model was the proximal 8 cm of the fibula in young adult dogs. In vascularized grafts, blood supply was re-established by microvascular re-anastomosis. Experimental controls were sham-operated and unoperated bones. Mid-graft test sections were subjected to loading-to-failure in torsion to determine the strength and stiffness. In both three- and six-month groups, vascularized grafts were significantly stronger and stiffer than contralateral nonvascularized grafts. Vascularized grafts were not significantly different from sham-operated bones. A 30-day regimen of cyclosporin A was found to have no measurable effect on mechanical properties for any individual treatment group. The results indicate that re-established blood supply can be a major factor in maintaining the mechanical integrity in large-segment cortical autografts.
Animals↗
Distal femoral replacement with allograft/prosthetic reconstruction for treatment of supracondylar fractures in patients with total knee arthroplasty.
Large-segment distal femoral allografts were used in conjunction with non-linked total knee prostheses to reconstruct bone deficits following supracondylar fracture of the femur in seven patients with previous total knee arthroplasties. Three patients with multiple medical problems died of unrelated causes prior to a minimum 2 year follow-up. Indications for surgery were previously failed attempts at osteosynthesis and significant fracture comminution, osteopenia, and intercondylar extension or femoral component loosening. Specifics of the surgical technique included subperiosteal excision of the involved distal femur with retention of a soft tissue sleeve containing the collateral ligaments and reconstruction with a large-segment allograft and a stemmed, semiconstrained total knee prosthesis. Cement fixation using pressurized technique with intramedullary plugging of the tibial and femoral canal was routinely used to secure the prosthesis/allograft construct to the host bone. Postoperative complications included one dislocation, which was successfully treated closed, and one popliteal artery injury, which was successfully repaired. There were no postoperative infections. Two patients, however, had some degree of persistent instability, warranting bracing at the time of last follow-up. Using the Knee Society rating system, the average knee score for these patients was 71, and the average pain score and function score were 33 and 49, respectively. Range of motion averaged 96 degrees. All of the femoral components were well fixed at last follow-up. Results of this study indicate that large-segment distal femoral allografts used in conjunction with nonlinked knee prostheses can be an acceptable method of treatment of these difficult reconstructive problems.
Aged↗
Characterization of cells with osteogenic potential from human marrow.
Studies using animal tissue suggest that bone marrow contains cells with the potential to differentiate into cartilage and bone. We report the extension of these studies to include human marrow. Bone marrow from male and female donors of various ages was obtained either from the femoral head or as aspirates from the iliac crest, and introduced into culture. Culture-adherent cells were expanded, subcultured, and then tested for bone and cartilage differentiation potential utilizing two different in vivo assays in nude mice. One assay involved subcutaneous implantation of porous calcium phosphate ceramics loaded with cultured, marrow-derived, mesenchymal cells; the other involved peritoneal implantation of diffusion chambers, also inoculated with cultured, marrow-derived, mesenchymal cells. Histological evaluation showed bone formation in ceramics implanted with cultured, marrow-derived, mesenchymal cells originating from both the femoral head and the iliac crest. Immunocytochemical analysis indicates that the bone is derived from the implanted human cells and not from the cells of the rodent host. No cartilage was observed in any of these ceramic grafts. In contrast, aliquots from the same preparations of cultured, marrow-derived, mesenchymal cells failed to form bone or cartilage in diffusion chambers. These data suggest that human marrow contains cells with osteogenic potential, which can be enriched and expanded in culture. Our findings also suggest that subcutaneous implantation of these cells in porous calcium phosphate ceramics may be a more sensitive in vivo assay than diffusion chambers for measuring their osteogenic lineage potential.
Animals↗
Use of an antibiotic impregnated polymethyl methacrylate intramedullary spacer for complicated revision total hip arthroplasty.
Revision total hip arthroplasty is frequently necessary in the presence of significant proximal femoral bone loss, periprosthetic fracture, or infection. In these situations, optimal reconstruction may sometimes warrant the use of special implants, including bone grafts. The emergent presentation of these cases or unexpected findings at the time of surgery can preclude the use of these treatment options. In cases of periprosthetic sepsis, delayed reimplantation may be the most successful approach to eradicate infection. In seven of these complicated revision total hip arthroplasties, the authors used an antibiotic-impregnated intramedullary polymethyl methacrylate spacer with delayed prosthetic reimplantation to allow for the use of these methods. Benefits of this technique include uncompromised radiographic evaluation of the proximal femur for design of a custom implant, if needed, stabilization of the proximal femur facilitating early mobilization of the patient in the case of periprosthetic fracture, and local delivery of antibiotics to the wound in the case of infection. The author's ability to reconstruct these total hip arthroplasties complicated by bone deficiency, fracture, and sepsis, was significantly improved with this use of this technique.
Adult↗
Culture-expanded periosteal-derived cells exhibit osteochondrogenic potential in porous calcium phosphate ceramics in vivo.
Chick tibial periosteal cells were enzymatically disaggregated, introduced into cell culture, and subcultured. These subcultured cells were combined with porous calcium phosphate ceramics and implanted into a subcutaneous site in athymic mice as an immunocompatible host to test the in vivo osteochondrogenic potential of this composite graft. These cells eventually gave rise to bone tissue in the pores of ceramics at the heterotopic implantation sites. The process of bone formation occurred through two different mechanisms: Intramembranous bone formation occurred at the peripheral pores of ceramics early, and endochondral bone formation occurred in the central pores later. Cultured chick muscle fibroblasts of the same-aged donor as controls did not form bone or cartilage under identical conditions to those of cultured periosteal-derived cells. These results raise the possibility that composite graft of cultured periosteal-derived cells and porous ceramics can be clinically used as a bone graft substitute in situations requiring bone augmentation or regeneration.
Animals↗
A small proteoglycan isolated from human cartilage containing a nonfunctional hyaluronic acid binding region.
A low buoyant density fraction (A4) was isolated from human cartilage by CsCl density gradient ultracentrifugation. This fraction contained a hydrodynamically small proteoglycan (Kav, 0.74 on Sepharose CL-2B) that reacted with monoclonal antibody 12/20/1C6 specific for the hyaluronic acid binding region (G1 globe) of the large aggregating high-density proteoglycan isolated from many animal cartilages. Despite the presence of the hyaluronic acid binding region, this small proteoglycan did not form proteoglycan aggregates with hyaluronan, not even in the presence of link protein.
Binding Sites↗
Culture-expanded human periosteal-derived cells exhibit osteochondral potential in vivo.
Periosteal cells were enzymatically liberated from human rib periostea obtained from autopsies of 37 donors with an age distribution ranging from 25 weeks of gestation to 88 years old. These cells were introduced into cell culture and subcultured when they reached confluence. After subculture, the adherent periosteal-derived cells showed a nondescript, fibroblast-like morphology in cell culture. The cells from various passages of each donor were tested for in vivo osteochondrogenic potential with three different assay methods in athymic mice: (a) inoculation assay--the cells were directly inoculated into a subcutaneous site, (b) porous ceramics assay--the cells were combined with porous calcium phosphate ceramics, and this composite graft was implanted into a subcutaneous site, and (c) diffusion chamber assay--the cells were loaded into diffusion chambers and cultured in the peritoneal cavity. Frozen-preserved and recultured periosteal-derived cells were also assayed in the same way. In cases of donors younger than 19 years old, cultured, periosteal-derived cells from up to several passages consistently formed bone and/or cartilage in each of the three assays. Frozen-preserved and recultured cells from these donors also formed bone and/or cartilage after introduction into the three in vivo assays. In cases of donors older than 22 years of age, cultured, periosteal-derived cells formed neither bone nor cartilage in vivo. Cultured muscle fibroblasts from some of the same donors did not form bone or cartilage when assayed in vivo under identical conditions. These results suggest that periosteal cells with osteochondrogenic potentials can be liberated from the periosteum of a rib of human donors up to a certain age. Importantly, this potential is retained after enzymatic liberation, cell culture, subculturing, and freeze preservation. The present results suggest that culture-expanded human periosteal-derived cells from young donors may be useful in the repair of skeletal defects to foster cell-mediated regeneration of skeletal tissues, and that this methodology can be used to elucidate cellular, molecular, and genetic disorders in various metabolic bone diseases and skeletal dysplasias.
Adult↗
Telemeterized in vivo hip joint force data: a report on two patients after total hip surgery.
Two telemeterized femoral components were implanted in two patients as part of normal total hip replacement procedures. The two components were instrumented to measure the three force components directed along: (a) the neck axis, (b) transverse to the neck axis and in the plane of the prosthesis, and (c) transverse to the neck axis and perpendicular to the plane of the prosthesis. Data were collected at multiple sessions during the early postoperative period for a number of standard activities, including gait, stair climbing, rising from a chair, single leg stance, double leg stance, ipsilateral and contralateral straight leg lifts while supine, ipsilateral flexion and extension while standing, and ipsilateral abduction while standing and lying on the contralateral side. These data are summarized and compared with the published results from analytic studies and with the results from previous studies using instrumented femoral components. Peak loads for gait during the period of study were roughly 2.7 body weights (BW) when the patients walked at their normal pace. Contact forces at the hip during stationary single leg stance approximated the peak loads during gait with values ranging from 2.1 to 2.8 BW. The highest forces recorded reached values approaching 5.5 BW and occurred during periods of instability while the patient engaged in stationary single leg stance. Our in vivo data indicate that forces generated during the above activities increase in magnitude quite rapidly during the early postoperative period and that during this period the patients have the ability to perform the activities of daily living without generating the high amplitude joint contact forces suggested by the results of dynamic studies. Joint contact forces during gait were found to depend on speed, but the high absolute magnitudes predicted by model studies were not supported by the in vivo data.
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Osteogenic potential of culture-expanded rat marrow cells as assayed in vivo with porous calcium phosphate ceramic.
It has been established that, when whole marrow is introduced into porous calcium phosphate ceramic, bone forms on the walls of the pores. To extend earlier studies, bone marrow cells derived from the femora of inbred rats were introduced into tissue culture and the adherent cells cultivated, mitotically expanded, passaged, harvested, placed in small cubes of porous calcium phosphate ceramics and grafted into subcutaneous sites of syngeneic rats. Marrow-derived, cultured mesenchymal cells introduced into ceramics showed strong osteogenic potential, with bone forming in the pore regions of ceramics as early as 2 wk after implantation. Osteogenesis could be observed after the eighteenth passage. With increasing passage number, the initiation of osteogenesis and the apparent rate of bone formation declined and the course of osteogenesis was delayed. In the future, it may be possible to culture marrow cells as a source for reparative cells for implantation back into autologous in vivo sites.
Animals↗
Bone mass and comparative rates of bone resorption and formation of fibular autografts: comparison of vascular and nonvascular grafts in dogs.
The metabolic fate of whole grafts that were either vascularized or nonvascularized were compared. This study was designed to quantify and correlate changes in bone resorption, formation, and mass in orthotopic, stably fixed, weight-bearing autografts. The grafts were 8-cm segments of the fibula that were internally fixed. Fibula segments subjected to sham operations, nonvascularized autografts, and vascularized autografts were studied in 16 dogs at three months after surgery. Three months prior to surgery the dogs were labeled repeatedly over two months with 3H-tetracycline and 3H-proline. Metabolic turnover of whole grafts was evaluated by quantifying loss of 3H-tetracycline for measuring postoperative resorption of bone mineral and loss of 3H-collagen for resorption of bone collagen. Net changes in bone dry weight, calcium, and collagen per whole grafts were obtained to determine differential changes in the mineral and matrix mass. The difference in change between bone resorption and bone mass was used to determine the amount of new bone formation that had replaced the resorbed bone. Vascularized autografts lost more mass (12%), and had more bone resorption (40%) and more bone formation (28%) than sham operated and unoperated fibulas. Nonvascularized grafts lost much more bone mass (48%) because resorption was large (61%) and formation was relatively small (13%). More new bone was formed in vascularized autografts than in nonvascularized autografts. During the incorporation of bone grafts, resorption is an early and rapid process, whereas formation is a late and slow process.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
Differential patterns of incorporation and remodeling among various types of bone grafts.
The early (3 months) and later (6 months) patterns of incorporation and bone formation have been evaluated histomorphometrically for different types of bone grafts; that is, vascularized and nonvascularized autografts with and without ciclosporin, and vascularized and nonvascularized dog leukocyte antigen (DLA)-mismatched allografts with and without ciclosporin. The vascularized bones were superior to the nonvascularized ones in healing and remodeling their grafted segments. In the autograft bones, ciclosporin did not alter the incorporation process 3 months after transplantation but delayed and increased the remodeling activities in the long run (6 months). Nonvascularized allografts underwent vigorous resorption, and were markedly porotic. Ciclosporin administration significantly reduced resorption and enhanced remodeling in nonvascularized allografts. The remodeling of allografts was similar to that of autografts in the presence of ciclosporin, but stopped soon after the administration of ciclosporin ceased.
Animals↗
The origin of bone formed in composite grafts of porous calcium phosphate ceramic loaded with marrow cells.
When porous calcium phosphate ceramic is combined with marrow cells and grafted either heterotopically or orthotopically, bone forms inside the pores on the surface of the ceramic beginning at three weeks after implantation. The question remains as to whether the newly formed bone is derived from host or donor cells. To study the origin of bone cells formed in these composite grafts of marrow cells and ceramic, quail marrow cells from long bones were introduced into ceramics and the composites were implanted into subcutaneous pouches of immunologically nonreactive athymic nude mice. The ceramics were recovered at two to 84 days following surgery, fixed, decalcified, embedded, sectioned, and examined for the location of a quail-specific nucleolar marker and the binding of a specific antiserum against quail cells. Our observations indicate that ceramic-associated osteogenesis is a biphasic phenomenon: an early phase, the first three to four weeks after implantation, in which donor cells are largely responsible for the observed osteogenesis, and a second phase, eight to 12 weeks postsurgery, in which host cell actions predominate. During the second stage, the ceramic pores begin to show the formation of marrow of host origin, and the mesenchymal marrow component appears to be osteogenic because the bone formed during this late postgrafting stage contains osteocytes of host and donor origin. The second phase therefore results in chimeric bone composed of quail and mouse. These studies clearly document the donor origin of the initial bone formation and indicate that marrow contains progenitor cells capable of forming de novo bone.
Animals↗
"Hybrid" total knee arthroplasty with the Miller-Galante prosthesis. A prospective clinical and roentgenographic evaluation.
Twenty-nine "hybrid" Miller-Galante total knee arthroplasties, in 22 patients, were evaluated prospectively and according to the clinical and roentgenographic guidelines of The Knee Society. Selection of this technique, incorporating an uncemented, porous-ingrowth femoral component and a cemented tibial component, was based on patient age, medical condition, activity level, and intraoperative assessment of bone quality and ligament competency. The average age of the patients at the time of surgery was 71 years. The average preoperative Knee Society Knee Score was 32; average pain score was 14; and the average function score, 47. After an average follow-up interval of 28 months postsurgery (minimum, 24 months), the average Knee Society Knee Score was 93; the average pain score, 47; and the average function score, 79. Range of motion averaged 110 degrees. Only one knee, with persistent pain of obscure origin, rated an unsuccessful result. No arthroplasties were revised for any reason. Twenty-three knees had fluoroscopically guided roentgenograms to assess the bone-prosthesis and bone-cement interfaces. No significant or progressive radiolucencies were noted under any of the components. No apparent adverse bone remodeling was associated with the uncemented femoral component. The fluoroscopically guided roentgenograms were significantly more sensitive in detecting interface radiolucencies than plain ones. Clinical and roentgenographic evidence of component loosening were absent in all patients. Results of this study suggest that the hybrid fixation technique can reliably provide satisfactory pain relief and restoration of function in properly selected patients. Potential advantages of an uncemented femoral component include decreased operative time, reduction of polyethylene wear from cement debris, and avoidance of a possible adverse biologic response to polymethylmethacrylate.
Aged↗
The osteogenic potential of culture-expanded rat marrow mesenchymal cells assayed in vivo in calcium phosphate ceramic blocks.
When whole marrow is introduced into porous calcium phosphate ceramic, bone forms on the walls of the pores. As an extension of earlier studies, bone marrow cells derived from the femora of inbred rats were introduced into tissue culture, and the adherent cells were cultivated, mitotically expanded, subcultured, harvested, placed in small cubes of porous calcium phosphate ceramic, and grafted into subcutaneous sites of syngeneic rats. Primary marrow-derived, cultured mesenchymal cells introduced into ceramic showed strong osteogenic potential, with bone forming in the pore regions of ceramic as early as two weeks after in vivo implantation; cartilage was observed infrequently in pores that appeared to be avascular. Osteogenesis could be observed after the 18th subculture (over 36 population doublings) when the cells were tested in ceramic at subcutaneous sites, whereas chondrogenesis was observed with only the first and second subcultured cells in the ceramic delivery vehicle. With increasing numbers of subcultures, the initiation of osteogenesis and the apparent rate of bone formation declined, and the course of osteogenesis was delayed. Cultured, marrow-derived mesenchymal cells, even after the 21st subculture (over 40 population doublings), exhibited a positive histochemical reaction for alkaline phosphatase. However, the in vivo osteogenic potential of these cells was not correlated with their alkaline phosphatase activity. The implantation of cell pellets or the injection of cell suspensions of fresh or cultured, adherent marrow cells never produced bone or cartilage in heterotopic sites. These data indicate that porous ceramic provides an excellent delivery vehicle for cells that are capable of osteogenic expression and suggest that the composite graft of marrow-derived mesenchymal cells and porous ceramic may be useful for repair of massive bone defects. It may be possible to culture marrow mesenchymal cells as a source for reparative cells for implantation back into autogeneic sites.
Alkaline Phosphatase↗
Ectopic induction of cartilage and bone by water-soluble proteins from bovine bone using a polyanhydride delivery vehicle.
Controlled release delivery vehicles for water-soluble osteogenic proteins from demineralized bovine bone matrix were constructed using polyanhydride polymers. The water-soluble proteins were isolated from a 4 M guanidine hydrochloride extract of bone matrix. The water-soluble proteins possessed Chondrogenic Stimulating Activity (CSA) when tested in stage 24 chick limb bud cell cultures, but were incapable of inducing cartilage or bone in vivo when implanted intramuscularly into mice by themselves. The polyanhydride polymers alone were also incapable of inducing ectopic cartilage or bone. However, when the water-soluble proteins were incorporated into the polymeric delivery vehicle, the combination was capable of inducing cartilage and bone up to 50% of the time. These results demonstrate that it is possible to use polyanhydride polymers as controlled-release delivery vehicles for soluble bioactive factors that interact with a local cell population.
Animals↗
Bone and cartilage formation in diffusion chambers by subcultured cells derived from the periosteum.
Periosteal cells were enzymatically isolated from the tibiae of young chicks, introduced into cell culture, allowed to reach confluence, and subcultured. The freshly isolated or subcultured cells were loaded into diffusion chambers and implanted into the peritoneal cavity of athymic mice to test their osteo-chondrogenic potential in a contained in vivo location. Freshly isolated periosteal cells formed both bone and cartilage tissue in such test chambers, but with a relatively low incidence. In contrast, cultured periosteal cells consistently gave rise to bone and cartilage even after 10 population doublings. With further passages of cells, the osteo-chondrogenic potential diminished substantially, until complete loss of expressivity at 16 population doublings or longer. Cultured muscle fibroblasts, when loaded into diffusion chambers under identical conditions to those of cultured periosteal cells, formed neither bone nor cartilage. These observations suggest that periosteal cells of young chicks contain subsets of progenitor cells or mesenchymal stem cells which possess the potential to differentiate into osteoblasts or chondrocytes, and this potential is retained after enzymatic isolation and for several population doublings in culture.
Animals↗