Chaoxu Liu

Influence of perfusion and compression on the proliferation and differentiation of bone mesenchymal stromal cells seeded on polyurethane scaffolds

In the present study, a porous meniscal-shaped scaffold consisting of polyurethane (PU)-based 1, 4-butanediisocyanate (BDI), which provided a 3-D culture condition for human bone mesenchymal stromal cells (hBMSC) was employed. A bioreactor was utilized to produce perfusion and mechanical stimulations. The viability, proliferation and fibro-cartilaginous differentiation of the hBMSC cultured on the PU-based meniscal scaffold were investigated during the perfusion and mechanical stimulation process. In addition, the mechanical properties of the cell-laden scaffolds were examined as well. Our finding indicated that the perfusion (10 ml/min) and on-off cyclic compressions mechanical stimulation (10% strain, 0.5 Hz, 4 times/day, 2 h/time with 4 h of rest thereafter) maintained the viability and promoted the proliferation of hBMSC over 2 weeks. The on-off cyclic compression caused a 1.85 fold increase in equilibrium modulus. Meanwhile, type I procollagen produced by hBMSC was increased for 3.02-fold after 2 weeks culture. On the other hand, the irrigating medium enhanced the synthesis of type III procollagen for 2.24-fold after 2 weeks. Tensile modulus was elevated for 2.02-fold in perfusion group after 1 week, which was decreased after 2 weeks unexpectedly. Our study suggests that the perfusion and on-off compression are promising to enhance the functional properties of the hBMSC-laden PU-based meniscal scaffold.

Meniscus reconstruction: today’s achievements and premises for the future

Injuries of the meniscus remain a burden for the development of premature cartilage degeneration and osteoarthritis. This review surveys all treatment options and focuses on the recent development of tissue engineering. Tissue engineering of the meniscus means a successful combination of cells, scaffolds and specific stimuli. Each element of the combination can be subject to variation. Studies investigating the optimum meniscus implant and previous steps in producing these implants are presented in this article. A comprehensive search of the English and German literature was performed in PubMed to retrieve appropriate manuscripts for review. Based on the literatures, autografts and allografts can delay the progress of osteoarthritis for a restricted time period, but several concerns persist. The biomechanical properties of the native meniscus are not copied entirely by the current existing autografts. Congruence, fixation, biocompatibility and potential infection will always remain as limitations for the users of allografts. Long-term results are still not available for meniscus prosthesis and even though it permits fast recovery, several aspects are questionable: bioincompatibility and a lack of cellular adhesion are likely to compromise their long-term fate. Currently, there is no ideal implant generated by means of tissue engineering. However, meniscus tissue engineering is a fast developing field, which promises to develop an implant that mimics histological and biomechanical properties of the native meniscus. At present several cell sources and scaffolds have been used successfully to grow 3-dimensional constructs. In future, optimal implants have to be developed using growth factors, modified scaffolds and stimuli that support cellular proliferation and differentiation to regenerate the native meniscus more closely.

The application of induced pluripotent stem cells for bone regeneration: current progress and prospects.

Loss of healthy bone tissue and dysosteogenesis are still common and significant problems in clinics. Cell-based therapy using mesenchymal stem cells (MSCs) has been performed in patients for quite some time, but the inherent drawbacks of these cells, such as the reductions in proliferation rate and osteogenic differentiation potential that occur with aging, greatly limit their further application. Moreover, embryonic stem cells (ESCs) have brought new hope to osteoregenerative medicine because of their full pluripotent differentiation potential and excellent performance in bone regeneration. However, the ethical issues involved in destroying human embryos and the immune reactions that occur after transplantation are two major stumbling blocks impeding the clinical application of ESCs. Instead, induced pluripotent stem cells (iPSCs), which are ESC-like pluripotent cells that are reprogrammed from adult somatic cells using defined transcription factors, are considered a more promising source of cells for regenerative medicine because they present no ethical or immunological issues. Here, we summarize the primary technologies for generating iPSCs and the biological properties of these cells, review the current advances in iPSC-based bone regeneration and, finally, discuss the remaining challenges associated with these cells, particularly safety issues and their potential application for osteoregenerative medicine.

Near-Infrared Spectroscopy Correlates with Established Histological Scores in a Miniature Pig Model of Cartilage Regeneration

Near-Infrared Spectroscopy (NIRS) could be of clinical relevance in modern cartilage regeneration.In a miniature pig model correlation of measurements and histologic scores have never been used before. The data analysis was part of an animal project that investigated the effects of seeding a chondrogenic and osteogenic scaffold with a bone-marrow-derived cell concentrate and reports the histological and mechanical properties. We created 20 osteochondral defects in the femoral condyles of 10 miniature pigs.The defects were left empty (E), filled with the grafted cylinder upside down (U), or with a combined scaffold (S) containing a spongy bone cylinder covered with a collagen membrane. In the fourth group, the same scaffolds were implanted but seeded with a stem cell concentrate (S+BMCC). The animals were euthanized after 3 months, and histologic and spectrometric analyses were performed. NIRS measurements were significantly higher in the central area of the defects of group S+BMCC compared to the central area of the defects of group U. In all groups, a correlation between NIRS and the histologic scores could be demonstrated though on different levels. In the central area, a good NIRS measurement correlates with low (good) histologic scores. In group E and group S, this negative correlation was significant (p=0.01). For the first time, NIRS was successfully used to evaluate osteochondral constructs in a miniature pig model.

The synergistic effect of bone forming peptide-1 and endothelial progenitor cells to promote vascularization of tissue engineered bone: Synergy of EPCs and BFP-1 in promoting vascularization

Large segmental bone defect repair remains a challenge in orthopedic surgeries. The tissue engineered bone graft will be a promising approach if vascularization of the graft is realized. In this study, beta-tricalcium phosphate (β-TCP) scaffold incorporated with bone forming peptide-1 (BFP-1) was fabricated. Endothelial progenitor cells (EPCs) were introduced as well. We investigated the effect of BFP-1 on the proliferation, differentiation, and angiogenic functions of EPCs. Additionally, segmental femur bone defect was created in rabbits. Prevascularized β-TCP scaffold was constructed and implanted into the bone defect. The vascularization and bone formation were evaluated after 4 and 12 weeks. The results showed that BFP-1 promoted the angiogenesis of EPCs through activating the activin receptor-like kinase-1/Smad pathway. The prevascularized tissue engineered bone graft enhanced capillary vessel in-growth and new bone formation. Significantly higher values of vascularization and radiographic grading scores were observed in groups involving EPCs and BFP-1, compared to β-TCP scaffold alone. In conclusion, the synergy between EPCs and BFP-1 improved the vascularization and new bone regeneration, which has great potentials in clinical applications.

The effect of anti-inflammatory and antifibrotic agents on fibroblasts obtained from arthrofibrotic tissue

Objectives The aims of this study were to determine whether the administration of anti-inflammatory and antifibrotic agents affect the proliferation, viability, and expression of markers involved in the fibrotic development of the fibroblasts obtained from arthrofibrotic tissue in vitro, and to evaluate the effect of the agents on arthrofibrosis prevention in vivo. Methods Dexamethasone, diclofenac, and decorin, in different concentrations, were employed to treat fibroblasts from arthrofibrotic tissue (AFib). Cell proliferation was measured by DNA quantitation, and viability was analyzed by Live/Dead staining. The levels of procollagen type I N-terminal propeptide (PINP) and procollagen type III N-terminal propeptide (PIIINP) were evaluated with enzyme-linked immunosorbent assay (ELISA) kits. In addition, the expressions of fibrotic markers were detected by real-time polymerase chain reaction (PCR). Fibroblasts isolated from healthy tissue (Fib) served as control. Further, a rabbit model of joint contracture was used to evaluate the antifibrotic effect of the three different agents. Results Dexamethasone maintained the viability and promoted the proliferation of AFib. Diclofenac decreased the viability and inhibited the cell proliferation during the first week of cultivation. However, decorin inhibited AFib proliferation and downregulated the expressions of fibrotic markers. Additionally, decorin could improve the flexion contracture angle and inhibit the deposition of interstitial matrix components in the rabbit joint model. Conclusion Decorin decreased the expression of myofibroblast markers in AFib, inhibited the proliferation of AFib, and prevented the initial procedure of arthrofibrosis in vivo, suggesting that decorin could be a promising treatment to inhibit the development of arthrofibrosis. Cite this article: X. Tang, S. Teng, M. Petri, C. Krettek, C. Liu, M. Jagodzinski. The effect of anti-inflammatory and antifibrotic agents on fibroblasts obtained from arthrofibrotic tissue: An in vitro and in vivo study. Bone Joint Res 2018;7:213–222. DOI: 10.1302/2046-3758.73.BJR-2017-0219.R2.

Influence of Hydrodynamic Pressure on the Proliferation and Osteogenic Differentiation of Bone Mesenchymal Stromal Cells Seeded on Polyurethane Scaffolds

Hydraulic pressure has recently been introduced as an effective stimulation in the field of tissue engineering. In this study, a polymer scaffold consisting of polyurethane (PU)-based 1, 4-butanediisocyanate was fabricated. A self-designed bioreactor was employed to produce perfusion and hydrodynamic pressure stimulations. The viability, proliferation and osteogenic differentiation of the rat bone mesenchymal stromal cell (rBMSC) growing in the polymer scaffold were investigated after hydrodynamic pressure stimulation. Additionally, the mechanical properties of the cell-laden constructs were also evaluated. Our findings suggested that the perfusion rate (10 mL/min) and low hydrodynamic pressure stimulation (60 mmHg, 0.5 Hz) maintained the viability of rBMSC during 2 weeks cultivation. The cell proliferation was promoted by 60 mmHg stimulation in the first week. The synthesis of alkaline phosphates and osteocalcin was enhanced after 2 weeks stimulation. Meanwhile, the equilibrium modulus of scaffold was increased by 1.85-fold using 60 mmHg hydrodynamic pressure stimulation. Additionally, type I and III procollagen produced by rBMSC was increased 4.92- and 3.02-fold, respectively. However, no encouraging results were detected in 120 mmHg hydrodynamic pressure group. Our study suggests that the 60 mmHg hydrodynamic pressure is a promising approach to enhance the functional properties of the rBMSC-laden PU-based bone scaffold.