P. Buma

Tissue ingrowth and degradation of two biodegradable porous polymers with different porosities and pore sizes.

Commonly, spontaneous repair of lesions in the avascular zone of the knee meniscus does not occur. By implanting a porous polymer scaffold in a knee meniscus defect, the lesion is connected with the abundantly vascularized knee capsule and healing can be realized. Ingrowth of fibrovascular tissue and thus healing capacity depended on porosity, pore sizes and compression modulus of the implant. To study the lesion healing potential, two series of porous polyurethanes based on 50/50 epsilon-caprolactone/L-lactide with different porosities and pore sizes were implanted subcutaneously in rats. Also, in vitro degradation of the polymer was evaluated. The porous polymers with the higher porosity, more interconnected macropores, and interconnecting micropores of at least 30 microm showed complete ingrowth of tissue before degradation had started. In implants with the lower macro-porosity and micropores of 10-15 microm degradation of the polymer occurred before ingrowth was completed. Directly after implantation and later during degradation of the polymer, PMN cells infiltrated the implant. In between these phases the foreign body reaction remained restricted to macrophages and giant cells. We can conclude that both foams seemed not suited for implantation in meniscal reconstruction while either full ingrowth of tissue was not realized before polymer degradation started or the compression modulus was too low. Therefore, foams must be developed with a higher compression modulus and more connections with sufficient diameter between the macropores.

Tissue engineering of the meniscus

Meniscus lesions are among the most frequent injuries in orthopaedic practice and they will inevitably lead to degeneration of the knee articular cartilage. The fibro-cartilage-like tissue of the meniscus is notorious for its limited regenerative capacity. Tissue engineering could offer new treatment modalities for repair of meniscus tears and eventually will enable the replacement of a whole meniscus by a tissue-engineered construct. Many questions remain to be answered before the final goal, a tissue-engineered meniscus is available for clinical implementation. These questions are related to the selection of an optimal cell type, the source of the cells, the need to use growth factor(s) and the type of scaffold that can be used for stimulation of differentiation of cells into tissues with optimal phenotypes. Particularly in a loaded, highly complex environment of the knee, optimal mechanical properties of such a scaffold seem to be of utmost importance. With respect to cells, autologous meniscus cells seems the optimal cell source for tissue engineering of meniscus tissue, but their availability is limited. Therefore research should be stimulated to investigate the suitability of other cell sources for the creation of meniscus tissue. Bone marrow stroma cells could be useful since it is well known that they can differentiate into bone and cartilage cells. With respect to growth factors, TGF-beta could be a suitable growth factor to stimulate cells into a fibroblastic phenotype but the problems of TGF-beta introduced into a joint environment should then be solved. Polyurethane scaffolds with optimal mechanical properties and with optimal interconnective macro-porosity have been shown to facilitate ingrowth and differentiation of tissue into fibro-cartilage. However, even these materials cannot prevent cartilage degeneration in animal models. Surface modification and/or seeding of cells into the scaffolds before implantation may offer a solution for this problem in the future.This review focuses on a number of specific questions; what is the status of the development of procedures for lesion healing and how far are we from replacing the entire meniscus by a (tissue-engineered) prosthesis. Subquestions related to the type of scaffold used are: is the degree of tissue ingrowth and differentiation related to the initial mechanical properties and if so, what is the influence of those properties on the subsequent remodelling of the tissue into fibro-cartilage; what is the ideal pore geometry and what is the optimal degradation period to allow biological remodelling of the tissue in the scaffold. Finally, we will finish with our latest results of the effect of tear reconstruction and the insertion of prostheses on articular cartilage degradation.

Culture of chondrocytes in alginate and collagen carrier gels

In this in vitro study, we compared the potential of collagen and alginate gels as carriers for chondrocyte transplantation and we studied the influence of demineralized bone matrix (DBM) on chondrocytes in the gels. Chondrocytes were assessed for cell viability, phenotype (histology), proliferation rate and sulfate incorporation. Collagen gels showed a significant increase in cell numbers, but the chondrocytes dedifferentiated into fibroblast-like cells from day 6 onwards. In alginate gels, initial cell loss was found, but the cells maintained their typical chondrocyte phenotype. Although the total quantity of proteoglycans initially synthesized per cell in collagen gel was significantly higher, expressed per cell, the quantity in alginate gel eventually surpassed collagen. No effects of culturing chondrocytes in combination with DBM could be demonstrated on cell proliferation and sulfate incorporation. The collagen and alginate gels have different advantages as carriers for chondrocyte transplantation. The high proliferation rate of chondrocytes in collagen gel may be an advantage, but the preservation of the chondrocyte phenotype and the gradually increasing proteoglycan synthesis in alginate gel is a promising method for creating a hyaline cartilage implant in vitro.

Crosslinked type II collagen matrices: preparation, characterization, and potential for cartilage engineering.

The limited intrinsic repair capacity of articular cartilage has stimulated continuing efforts to develop tissue engineered analogues. Matrices composed of type II collagen and chondroitin sulfate (CS), the major constituents of hyaline cartilage, may create an appropriate environment for the generation of cartilage-like tissue. In this study, we prepared, characterized, and evaluated type 11 collagen matrices with and without CS. Type II collagen matrices were prepared using purified, pepsin-treated, type II collagen. Techniques applied to prepare type I collagen matrices were found unsuitable for type II collagen. Crosslinking of collagen and covalent attachment of CS was performed using 1-ethyl-3-(3-dimethyl aminopropyl)carbodiimide. Porous matrices were prepared by freezing and lyophilization, and their physico-chemical characteristics (degree of crosslinking, denaturing temperature, collagenase-resistance, amount of CS incorporated) established. Matrices were evaluated for their capacity to sustain chondrocyte proliferation and differentiation in vitro. After 7 d of culture, chondrocytes were mainly located at the periphery of the matrices. In contrast to type I collagen, type II collagen supported the distribution of cells throughout the matrix. After 14 d of culture, matrices were surfaced with a cartilagenous-like layer, and occasionally clusters of chondrocytes were present inside the matrix. Chondrocytes proliferated and differentiated as indicated by biochemical analyses, ultrastructural observations, and reverse transcriptase PCR for collagen types I, II and X. No major differences were observed with respect to the presence or absence of CS in the matrices.

Resurfacing potential of heterologous chondrocytes suspended in fibrin glue in large full-thickness defects of femoral articular cartilage : an experimental study in the goat

A large full-thickness articular-cartilage defect was created in the medial femoral condyle of 32 adult goats. The defects were xenografted with isolated rabbit chondrocytes suspended in fibrin glue. Sham operated goats, where only a standardized defect was created, were used as controls. Results of cartilage repair were assessed after 3, 8, 13, 26 and 52 weeks. The repair tissue was evaluated macroscopically, histologically and biochemically. Results indicated that xenografted rabbit chondrocytes survived the transplantation and maintained their potential to produce matrix in fibrin glue, particularly if they were located in a non-weight-bearing area. In terms of an immunological reaction to xenografted chondrocytes, only mild signs of synovitis were observed in both groups and rejection of transplanted cells did not occur. From 3 weeks gradually progressive resolvement of the fibrin glue was observed with subsequent replacement by fibrous tissue. Initially xenografted defects histologically showed better tendency for cartilage regeneration, however, 52 weeks after surgery no significant differences could be detected in the repair tissue of both groups macroscopically, histologically and on biochemical scoring. The amount of collagen type II in the newly synthesized matrix was 75% 1 year after surgery. This study shows that isolated heterologous chondrocytes can be used for transplantation in articular cartilage defects, however, fibrin glue does not offer enough biomechanical support to the cells to maintain its function as a three-dimensional scaffold.

Acetabular reconstruction with impacted morsellised cancellous bone graft and cement: A 10- to 15-year follow-up of 60 revision arthroplasties

We report a long-term review of 60 acetabular components revised using impacted, morsellised bone allografts and a cemented polyethylene cup. The acetabular defects were cavitary (37) or combined (23). Follow-up was for a mean 11.8 years (10 to 15). Further revision was needed in five hips, two for septic and three for aseptic loosening. The overall survival rate at 11.8 years was 90%; excluding the septic cases it was 94%. Acetabular reconstruction with impacted morsellised cancellous grafts and cement gives satisfactory long-term results.

Incorporation of morselized bone grafts: a study of 24 acetabular biopsy specimens.

Animal studies have shown almost complete incorporation of impacted morselized bone grafts. To determine whether this also is true in humans, 24 acetabular bone biopsy specimens from 21 hips of 20 patients were examined. Biopsy specimens were obtained 3 months to 15 years after acetabular reconstruction in primary and revision total hip arthroplasties in combination with a cemented cup. Histologic examination showed rapid revascularization of the graft, directly followed by osteoclastic resorption and woven bone formation on the graft remnants. New bone also was formed on fibrin accumulations or without any scaffold in the fibrous stroma tissue that had invaded the graft. Thereafter the mixture of graft, new bone, and fibrin was remodeled completely into a new trabecular structure, with normal lamellar bone and only scarce remnants of graft material. Localized areas of nonincorporated bone graft surrounded by fibrous tissue remained, irrespective of the followup period. Large nonincorporated fragments of cartilage also were found, particularly in cases in which femoral head bone chips were produced by a bone mill. In general, impacted trabecular bone chips incorporate by a mechanism that is similar to that previously observed in animal studies. In some patients, however, areas of nonincorporated bone graft remained and long-term alterations were found, probably related to the loosening process.

Acetabular reconstruction with bone impaction grafting and a cemented cup: 20 years' experience.

Acetabular bone stock loss compromises the outcome in primary and revision total hip arthroplasty. In 1979, a biologic method was introduced with tightly impacted cancellous allografts in combination with a cemented polyethylene cup for acetabular reconstruction. With this technique, it is possible to replace the loss of bone and to repair normal hip mechanics and hip function with a standard implant. Based on the authors’ 20 years experience, a review of the long-term data is presented in primary total hip arthroplasty with preexisting acetabular bone stock loss, primary total hip arthroplasty in rheumatoid arthritis, patients who had bone impaction when younger than 50 years, and in acetabular revisions. The survival rate with revision of the cup for aseptic loosening as the end point was 94% at 10 to 17 years, 90% at 10 to 18 years, 91% at 10 to 17 years, and 92% at 10 to 15 years. From biopsy specimens from humans and histologic data in animal experiments the incorporation of these impacted bone chips was proven. The acetabular bone impaction technique using large morselized bone chips (range, 0.7–1 cm) and a cemented cup is a reliable technique with favorable long-term outcome.

Morsellized allografts for fixation of the hip prosthesis femoral component. A mechanical and histological study in the goat

To simulate femoral intramedullary bone stock loss in revision surgery of failed total hip arthroplasties, a method was developed using impacted trabecular bone grafts. In 14 goats a cemented total hip arthroplasty was performed, fixating the stem within a circumferential construction of bone allografts. After 6 or 12 weeks, 4 goats were used for mechanical tests and 3 for histology. The stability of the stems was determined in a loading experiment with roentgen-stereophotogrammetric analysis; loads of up to 1.44 times body weight were used. One aseptic loosening was seen with gross movements. In the other cases the most important movements were axial rotations (max. 0.24 degrees under 800 N) and axial translations (max. 0.16 mm under 800 N). After unloading some elastic recovery occurred. There were no differences between the 6 and 12-week groups. Histologically, revascularization and remodeling of the grafts were evident. Bone apposition and bone resorption of the grafts resulted in a mixture of graft and new bone. There was more new bone formation in the 12-week group, but the process was not yet completed. The use of impacted trabecular bone grafts in cases of severe intramedullary bone stock loss seems to be a promising revision technique.

Impacted graft incorporation after cemented acetabular revision. Histological evaluation in 8 patients.

We took core biopsies from the acetabulum in 8 patients (at reoperation) after a previous revision with impacted cancellous allograft chips in combination with cement. Except for one biopsy specimen, the graft showed different stages of incorporation. In the specimens taken at 4 months, revascularization of the graft was found. Osteoclasts had removed parts of the graft, while woven bone had formed on the remnants of the graft and in the stroma that was invading the graft. Subsequent specimens showed that this mixture of graft and new bone was in due time remodeled into a normal trabecular bony structure with viable bone marrow that contained little or no remnants of the original graft. The graft-cement interface was present in 4 biopsies taken at 1, 22, 28, and 72 months. The specimen obtained 28 months after revision showed vital bone locally in direct contact with the cement layer; however, a soft tissue interface predominated.