Condyles Of Rabbits Research Articles

Compositional Variations in Calcium Phosphate Cement and Poly(Lactic-Co-Glycolic-Acid) Porogens Do Not Affect the Orthotopic Performance of Calcium Phosphate Cement/Poly(Lactic-Co-Glycolic-Acid) Cements.

Calcium phosphate cement (CPC) has evolved as an appealing bone substitute material, especially since CPCs were combined with poly(lactic-co-glycolic acid) (PLGA) porogens to render the resulting CPC/PLGA composite degradable. In view of the multiple variables of CPC and PLGA used previously, the effect of CPC composition and PLGA porogen morphology (i.e., microspheres versus microparticles) on the biological performance of CPC/PLGA has not yet been investigated. Consequently, we here aimed to evaluate comparatively various CPC/PLGA formulations varying in CPC composition and PLGA porogen morphology on their performance in a rabbit femoral condyle bone defect model. CPCs with a composition of 85 wt% α-TCP, 15 wt% dicalcium phosphate anhydrate (DCPA) and 5 wt% precipitated hydroxyapatite (pHA), or 100 wt% α-TCP were combined with spherical or irregularly shaped PLGA porogens (CPC/PLGA ratio of 60:40 wt% for all formulations). All CPC/PLGA formulations were applied via injection in bone defects, as created in the femoral condyle of rabbits, and retrieved for histological evaluation after 6 and 12 weeks of implantation. Descriptive histology and quantitative histomorphometry (i.e., material degradation and new bone formation) were used for analyses. Descriptively, all CPC/PLGA formulations showed material degradation at the periphery of the cement within 6 weeks of implantation. After 12 weeks, bone formation was observed extending into the defect core, replacing the degraded CPC/PLGA material. Quantitatively, similar material degradation (up to 87%) and new bone formation (up to 28%) values were observed, irrespective of compositional variations of CPC/PLGA formulations. These data prove that neither the CPC compositions nor the PLGA porogen morphologies as used in this work affect the biological performance of CPC/PLGA formulations in a rabbit femoral condyle bone defect model.

Dissolving magnesium hydroxide implants enhance mainly cancellous bone formation whereas degrading RS66 implants lead to prominent periosteal bone formation in rabbits

Bone fractures often require internal fixation using plates or screws. Normally, these devices are made of permanent metals like titanium providing necessary strength and biocompatibility. However, they can also cause long-term complications and may require removal. An interesting alternative are biocompatible degradable devices, which provide sufficient initial strength and then degrade gradually. Among other materials, biodegradable magnesium alloys have been developed for craniofacial and orthopaedic applications. Previously, we tested implants made of magnesium hydroxide and RS66, a strong and ductile ZK60-based alloy, with respect to biocompatibility and degradation behaviour. Here, we compare the effects of dissolving magnesium hydroxide and RS66 cylinders on bone regeneration and bone growth in rabbit condyles using microtomographical and histological analysis. Both magnesium hydroxide and RS66 induced a considerable osteoblastic activity leading to distinct but different spatio-temporal patterns of cancellous and periosteal bone growth. Dissolving RS66 implants induced a prominent periosteal bone formation on the medial surface of the original condyle whereas dissolving magnesium hydroxide implants enhance mainly cancellous bone formation. Especially periosteal bone formation was completed after 6 and 8 weeks, respectively. The observed bone promoting functions are in line with previous reports of magnesium stimulating cancellous and periosteal bone growth and possible underlying signalling mechanisms are discussed. Statement of significanceBiodegradable magnesium based implants are promising candidates for use in orthopedic and traumatic surgery. Although these implants are in the scientific focus for a long time, comparatively little is known about the interactions between degrading magnesium and the biological environment. In this work, we investigated the effects of two degrading cylindrical magnesium implants (MgOH2 and RS66) both on bone regeneration and on bone growth. Both MgOH2 and RS66 induce remarkable osteoblastic activities, however with different spatio-temporal patterns regarding cancellous and periosteal bone growth. We hypothesize that degradation products do not diffuse directionless away, but are transported by the restored blood flow in specific spatial patterns which is also dependent on the used surgical technique.

Enhancing osteoblast proliferation and bone regeneration by poly (amino acid)/selenium-doped hydroxyapatite

Among various biomaterials employed for bone repair, composites with good biocompatibility and osteogenic ability had received increasing attention from biomedical applications. In this study, we doped selenium (Se) into hydroxyapatite (Se-HA) by the precipitation method, and prepared different amounts of Se-HA-loaded poly (amino acid)/Se-HA (PAA/Se-HA) composites (0, 10 wt%, 20 wt%, 30 wt%) by in-situ melting polycondensation. The physical and chemical properties of PAA/Se-HA composites were characterized by x-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), scanning electron microscopy (SEM) and their mechanical properties. XRD and FT-IR results showed that PAA/Se-HA composites contained characteristic peaks of PAA and Se-HA with amide linkage and HA structures. DSC and TGA results specified the PAA/Se-HA30 composite crystallization, melting, and maximum weight loss temperatures at 203.33 °C, 162.54 °C, and 468.92 °C, respectively, which implied good thermal stability. SEM results showed that Se-HA was uniformly dispersed in PAA. The mechanical properties of PAA/Se-HA30 composites included bending, compressive, and yield strengths at 83.07 ± 0.57, 106.56 ± 0.46, and 99.17 ± 1.11 MPa, respectively. The cellular responses of PAA/Se-HA composites in vitro were studied using bone marrow mesenchymal stem cells (BMSCs) by cell counting kit-8 assay, and results showed that PAA/Se-HA30 composites significantly promoted the proliferation of BMSCs at the concentration of 2 mg ml−1. The alkaline phosphatase activity (ALP) and alizarin red staining results showed that the introduction of Se-HA into PAA enhanced ALP activity and formation of calcium nodule. Western blotting and Real-time polymerase chain reaction results showed that the introduction of Se-HA into PAA could promoted the expression of osteogenic-related proteins and mRNA (integrin-binding sialoprotein, osteopontin, runt-related transcription factor 2 and Osterix) in BMSCs. A muscle defect at the back and a bone defect at the femoral condyle of New Zealand white rabbits were introduced for evaluating the enhancement of bone regeneration of PAA and PAA/Se-HA30 composites. The implantation of muscle tissue revealed good biocompatibility of PAA and PAA/Se-HA30 composites. The implantation of bone defect showed that PAA/Se-HA30 composites enhanced bone formation at the defect site (8 weeks), exhibiting good bone conductivity. Therefore, the PAA-based composite was a promising candidate material for bone tissue regeneration.

Continuous Low-Intensity Ultrasound Improves Cartilage Repair in Rabbit Model of Subchondral Injury.

Subchondral drilling (SD), a bone marrow stimulation technique, is used to repair cartilage lesions that lack regenerative potential. Cartilage repair outcomes upon SD are typically fibrocartilaginous in nature with inferior functionality. The lack of cues to foster the chondrogenic differentiation of egressed mesenchymal stromal cells upon SD can be attributed for the poor outcomes. Continuous low-intensity ultrasound (cLIUS) at 3.8 MHz is proposed as a treatment modality for improving cartilage repair outcomes upon marrow stimulation. Bilateral defects were created by SD on the femoral medial condyle of female New Zealand white rabbits (n = 12), and the left joint received cLIUS treatment (3.8 MHz, 3.5 Vpp, 8 min/application/day) and the contralateral right joint served as the control. On day 7 postsurgery, synovial fluid was aspirated, and the cytokine levels were assessed by Quantibody™ assay. Rabbits were euthanized at 8 weeks and outcomes were assessed macroscopically and histologically. Defect areas in the right joints exhibited boundaries, incomplete fill, irregular cartilage surfaces, loss of glycosaminoglycan (GAG), and absence of chondrocytes. In contrast, the repaired defect area in the joints that received cLIUS showed complete fill, positive staining for GAG with rounded chondrocyte morphology, COL2A1 staining, and columnar organization. Synovial fluid collected from cLIUS-treated left knee joints had lower levels of IL1, TNFα, and IFNγ when compared to untreated right knee joints, alluding to the potential of cLIUS to mitigate early inflammation. Further at 8 weeks, left knee joints (n = 12) consistently scored higher on the O'Driscoll scale, with a higher percent hyaline cartilage score. No adverse impact on bone or change in the joint space was noted. Upon a single exposure of cLIUS to TNFα-treated cells, nuclear localization of pNFκB and SOX9 was visualized by double immunofluorescence and the expression of markers associated with the NFκB pathway was assayed by quantitative real-time polymerase chain reaction. cLIUS extends its chondroprotective effects by titrating pNFκB levels, preventing its nuclear translocation, while maintaining the expression of SOX9, the collagen II transcription factor. Our combined results demonstrate that healing of chondral defects treated with marrow stimulation by SD can be accelerated by employing cLIUS regimen that possesses chondroinductive and chondroprotective properties. Impact statement Repair of cartilage represents an unsolved biomedical burden. In vitro, continuous low-intensity ultrasound (cLIUS) has been demonstrated to possess chondroinductive and chondroprotective potential. To our best knowledge, the use of cLIUS to improve cartilage repair outcomes upon marrow stimulation, in vivo, has not been reported and our work reported here fills that gap. Our results demonstrated enhanced cartilage repair outcomes under cLIUS (3.8 MHz) in a rabbit model of subchondral injury by subchondral drilling. Enhanced repair stemmed from mesenchymal stem cell differentiation in vivo and the subsequent synthesis of articular cartilage-specific matrix.

Simvastatin/hydrogel-loaded 3D-printed titanium alloy scaffolds suppress osteosarcoma via TF/NOX2-associated ferroptosis while repairing bone defects

Postoperative anatomical reconstruction and prevention of local recurrence after tumor resection are two vital clinical challenges in osteosarcoma treatment. A three-dimensional (3D)-printed porous Ti6Al4V scaffold (3DTi) is an ideal material for reconstructing critical bone defects with numerous advantages over traditional implants, including a lower elasticity modulus, stronger bone-implant interlock, and larger drug-loading space. Simvastatin is a multitarget drug with anti-tumor and osteogenic potential; however, its efficiency is unsatisfactory when delivered systematically. Here, simvastatin was loaded into a 3DTi using a thermosensitive poly (lactic-co-glycolic) acid (PLGA)-polyethylene glycol (PEG)-PLGA hydrogel as a carrier to exert anti-osteosarcoma and osteogenic effects. Newly constructed simvastatin/hydrogel-loaded 3DTi (Sim-3DTi) was comprehensively appraised, and its newfound anti-osteosarcoma mechanism was explained. Specifically, in a bone defect model of rabbit condyles, Sim-3DTi exhibited enhanced osteogenesis, bone in-growth, and osseointegration compared with 3DTi alone, with greater bone morphogenetic protein 2 expression. In our nude mice model, simvastatin loading reduced tumor volume by 59%–77 % without organic damage, implying good anti-osteosarcoma activity and biosafety. Furthermore, Sim-3DTi induced ferroptosis by upregulating transferrin and nicotinamide adenine dinucleotide phosphate oxidase 2 levels in osteosarcoma both in vivo and in vitro. Sim-3DTi is a promising osteogenic bone substitute for osteosarcoma-related bone defects, with a ferroptosis-mediated anti-osteosarcoma effect.

In Vitro and In Vivo Bone Regeneration Assessment of Titanium-Doped Waste Eggshell-Derived Hydroxyapatite in the Animal Model.

In this work, a titanium-doped hydroxyapatite (HAp) scaffold was produced from two different sources (natural eggshell and laboratory-grade reagents) to compare the efficacy of natural and synthetic resources of HAp materials on new bone regeneration. This comparative study also reports the effect of Ti doping on the physical, mechanical, and in vitro as well as in vivo biological properties of the HAp scaffold. Pellets were prepared in the conventional powder metallurgy route, compacted, and sintered at 900 °C, showing sufficient porosity for bony ingrowth. The physical-mechanical characterizations were performed by density, porosity evaluation, XRD, FTIR, SEM analysis, and hardness measurement. In vitro interactions were evaluated by bactericidal assay, hemolysis, MTT assay, and interaction with simulated body fluid. All categories of pellets showed absolute nonhemolytic and nontoxic character. Furthermore, significant apatite formation was observed on the Ti-doped HAp samples in the simulated body fluid immersion study. The developed porous pellets were implanted to assess the bone defect healing in the femoral condyle of healthy rabbits. A 2 month study after implantation showed no marked inflammatory reaction for any samples. Radiological analysis, histological analysis, SEM analysis, and oxytetracycline labeling studies depicted better invasion of mature osseous tissue in the pores of doped eggshell-derived HAp scaffolds as compared to the undoped HAp, and laboratory-made samples. Quantification using oxytetracycline labeling depicted 59.31 ± 1.89% new bone formation for Ti-doped eggshell HAp as compared to Ti-doped pure HAp (54.41 ± 1.93) and other undoped samples. Histological studies showed the presence of abundant osteoblastic and osteoclastic cells in Ti-doped eggshell HAp in contrast to other samples. Radiological and SEM data also showed similar results. The results indicated that Ti-doped biosourced HAp samples have good biocompatibility, new bone-forming ability, and could be used as a bone grafting material in orthopedic surgery.

Degradation and Bone-Contact Biocompatibility of Two Drillable Magnesium Phosphate Bone Cements in an In Vivo Rabbit Bone Defect Model.

The use of bone-cement-enforced osteosynthesis is a growing topic in trauma surgery. In this context, drillability is a desirable feature for cements that can improve fracture stability, which most of the available cement systems lack. Therefore, in this study, we evaluated a resorbable and drillable magnesium-phosphate (MgP)-based cement paste considering degradation behavior and biocompatibility in vivo. Two different magnesium-phosphate-based cement (MPC) pastes with different amounts of phytic acid (IP 6) as setting retarder (MPC 22.5 and MPC 25) were implanted in an orthotopic defect model of the lateral femoral condyle of New Zealand white rabbits for 6 weeks. After explantation, their resorption behavior and material characteristics were evaluated by means of X-ray diffraction (XRD), porosimetry measurement, histological staining, peripheral quantitative computed tomography (pQCT), cone-beam computed tomography (CBCT) and biomechanical load-to-failure tests. Both cement pastes displayed comparable results in mechanical strength and resorption kinetics. Bone-contact biocompatibility was excellent without any signs of inflammation. Initial resorption and bone remodeling could be observed. MPC pastes with IP 6 as setting retardant have the potential to be a valuable alternative in distinct fracture patterns. Drillability, promising resorption potential and high mechanical strength confirm their suitability for use in clinical routine.

Icariin-conditioned serum combined with chitosan attenuates cartilage injury in rabbit knees with osteochondral defect

BackgroundKnee osteoarthritis (KOA) is one of the most common degenerative diseases. Its development is closely related to cartilage injury and subchondral bone remodeling homeostasis. In the present study, we combined icariin-conditioned serum (ICS) with thiolated chitosan (CSSH), a material widely used in tissue engineering for cartilage repair, to demonstrate its effect on the repair of cartilage damage and abnormal subchondral remodeling.MethodsNew Zealand rabbits were undergoing surgery for cartilage defect, and joint cavity injection was performed in each group with 0.5 mL normal saline (NS), ICS, CSSH and ICS-CSSH in the right joint every week for five times. Positioning performance was observed using VICON motion capture system. Glycosaminoglycans (GAG) secretion of articular fluid was assessed. Osteoarthritis Research Society International (OARSI) score and immunohistochemical (IHC) analysis including H&E, Safranin O and collagen II staining were employed to evaluate the morphologic repair of cartilage and subchondral bone. mRNA expression of COL2A1, MMP13 and ADAMTS5 was detected in chondrocytes from injury area.ResultsICS combined with CSSH attenuated cartilage injury and abnormal subchondral remodeling in rabbits with KOA. ICS and CSSH groups showed slight improvement in positioning performance, while ICS-CSSH group exhibited better positioning performance. ICS-CSSH group showed increased GAG secretion of articular fluid and expression of COL2A1 in articular chondrocytes. Furthermore, both macroscopic observation and IHC analysis showed femoral condyle in ICS-CSSH rabbits were repaired with more native cartilage and subchondral bone regeneration.ConclusionsICS combined with CSSH could promote the repair of osteochondral defect and stabilize subchondral bone remodeling in rabbit knees.

Bioactive graphene oxide-functionalized self-expandable hydrophilic and osteogenic nanocomposite for orthopaedic applications.

Polymethyl methacrylate (PMMA) bone cement (PBC) is commonly used in orthopaedic surgery. However, polymerization volumetric shrinkage, exothermic injury, and low bioactivity prevent PBC from being an ideal material. The developed expandable P(MMA-AA-St) well overcomes the volumetric shrinkage of PBC. However, its biomechanical properties are unsatisfactory. Herein, graphene oxide (GO), a hydrophilic material with favourable biomechanics and osteogenic capability, was added to P(MMA-AA-St) to optimize its biomechanics and bioactivity. The GO-modified self-expandable P(MMA-AA-St)-GO nanocomposite (PGBCs) exhibited outstanding compressive strength (>70​MPa), water absorption, and volume expansion, as well as a longer handling time and a reduced setting temperature. The cytocompatibility of PGBCs was superior to that of PBC, as demonstrated by CCK-8 assay, live-dead cell staining, and flow cytometry. In addition, better osteoblast attachment was observed, which could be attributed to the effects of GO. The improved level of osteogenic gene and protein expression further illustrated the improved cell-material interactions between osteoblasts and PGBCs. The results of an in vivo study performed by filling bone defects in the femoral condyles of rabbits with PGBCs demonstrated promising intraoperative handling properties and convenient implantation. Blood testing and histological staining demonstrated satisfactory in vivo biosafety. Furthermore, bone morphological and microarchitecture analyses using bone tissue staining and micro-CT scanning revealed better bone-PGBCs contact and osteogenic capability. The results of this study indicate that GO modification improved the physiochemical properties, cytocompatibility, and osteogenic capability of P(MMA-AA-St) and overcame the drawbacks of PBC, allowing its material derivatives to serve as effective implantable biomaterials.

Comparison of bone ingrowth between two porous titanium alloy rods with biogenic lamellar structures and diamond crystal lattice on femoral condyles in rabbits

PurposeThe comparison of bone ingrowth between two types of porous titanium alloy rods with different micro-architectures including diamond crystal lattice (Re-rod) and biogenic lamellar configurations (Bi-rod) on femoral condyles was investigated in this study. MethodsTwelve rabbits were used. Re-rod (Re-rod group) and Bi-rod (Bi-rod group) were implanted randomly in femoral condyles of each rabbits respectively. Bone ingrowth of these two rods were investigated and compared. 4 and 12 weeks after the operation, X-ray, micro-CT and histological examinations were performed. ResultsNo femoral condyle fracture and rod defluxion in the two groups was noted in the X-ray images during the observation period. Micro-CT images showed that all metal trabeculae in the Bi-rod group were covered by new bone at 4 and 12 weeks, whereas partial metal trabeculae in the Re-rod group were still uncovered at 12 weeks. Histological images showed that there was new bone growth in the centre and periphery of Bi-rods at 4 and 12 weeks, and there were several areas without new bone ingrowth at 4 and 12 weeks in the centre of Re-rods. In micro-CT analysis, the bone volume to total volume (BV/TV) of the volume of interest (VOI) of the Bi-rod group was higher than in the Re-rod group [(0.0794 ± 0.0021) % Vs (0.0521 ± 0.0032) % and (0.0875 ± 0.0039) % Vs (0.0702 ± 0.0028) % respectively, P < 0.05] at 4 weeks and 12 weeks. Whereas, the mean trabecular thickness (Tb.Th) values of VOI between the two groups were not significantly statistically different at 4 and 12 weeks. In histological analysis, the BV/TV of the VOI of the Bi-rod group was higher than in the Re-rod group [(0.0624 ± 0.0021) % Vs (0.0435 ± 0.0028) % and (0.0675 ± 0.0024) % Vs (0.0476 ± 0.0031) % respectively, P < 0.05] at 4 weeks and 12 weeks. ConclusionThese results showed that Bi-rods got better bone ingrowth in femoral condyles of rabbits compared with Re-rods.

A Preclinical Study of Bone Marrow Buffy Coat Components and Cartilage Regeneration

Background: Autologous bone marrow buffy coat transplantation possesses obvious advantages in the therapy of cartilage defects. However, there is no definite research on the specific effective components of bone marrow buffy coat and their mechanism of cartilage regeneration. Moreover, bone marrow buffy coat is difficult to fix onto the damaged cartilage area. We evaluated the composition of Buffy coat and the combined use of hyaluronic acid (HA) as a scaffold material for repairing cartilage defects. Methods and Materials: We extracted the bone marrow from the anterior superior iliac crest of the rabbit, centrifuged it to obtain buffy coat, and analyzed the components of buffy coat by enzyme-linked immunosorbent assay (ELISA). Buffy coat+fibrinogen/HA group, mesenchymal stem cell (MSC)+fibrinogen/HA group, MSC+ Transforming Growth Factorβ(TGF-β)+ fibrinogen/HA group were culturedin vitroand observed by staining. In addition, we made damage to the femoral condyle of rabbits and divided them into groups: HA group, buffy coat group, buffy coat with HA group. Each group was assessed for cartilage regeneration by visual observation, histological at 4 weeks and 8 weeks, and biochemical analysis at 8 weeks postoperatively. One-way ANOVA and LSD were used for statistical analysis. Results: Buffy coat have a variety of growth factors, inflammatory factors, and anti-inflammatory factors that stimulate the MSCs’ regeneration. Buffy coat can differentiate into cartilage without TGF-βstimulationin vitro. The cartilage regeneration ability of buffy coat and buffy coat+HA is strong, and the combination of buffy coat and gel scaffold HA can make cartilage formation ability more stablein vivo. Conclusion: MSC and cytokines in buffy coat synergistically promote cartilage regeneration. Gel scaffold HA enhances the effect of buffy coat on cartilage attachment and regeneration of cartilage defects.

Type I collagen scaffold with WNT5A plasmid for in situ cartilage tissue engineering.

Cartilage tissue lacks the ability to heal. Cartilage tissue engineering using cell-free scaffolds has been increasingly used in recent years. This study describes the use of a type I collagen scaffold combined with WNT5A plasmid to promote chondrocyte proliferation and differentiation in a rabbit osteochondral defect model. Type I collagen was extracted and fabricated into a collagen scaffold. To improve gene transfection efficiency, a cationic chitosan derivative N,N,N-trimethyl chitosan chloride (TMC) vector was used. A solution of TMC/WNT5A complexes was adsorbed to the collagen scaffold to prepare a WNT5A scaffold. Osteochondral defects were created in the femoral condyles of rabbits. The rabbits were divided into defect, scaffold, and scaffold with WNT5A groups. At 6 and 12 weeks after creation of the osteochondral defects, samples were collected from all groups for macroscopic observation and gene expression analysis. Samples from the defect group exhibited incomplete cartilage repair, while those from the scaffold and scaffold with WNT5A groups exhibited "preliminary cartilage" covering the defect. Cartilage regeneration was superior in the scaffold with WNT5A group compared to the scaffold group. Safranin O staining revealed more proteoglycans in the scaffold and scaffold with WNT5A groups compared to the defect group. The expression levels of aggrecan, collagen type II, and SOX9 genes were significantly higher in the scaffold with WNT5A group compared to the other two groups. Type I collagen scaffold showed effective adsorption and guided the three-dimensional arrangement of stem cells. WNT5A plasmid promoted cartilage repair by stimulating the expression of aggrecan, type II collagen, and SOX9 genes and proteins, as well as inhibiting cartilage hypertrophy.

Location-Specific Study of Young Rabbit Femoral Cartilage by Quantitative µMRI and Polarized Light Microscopy.

ObjectiveMicroscopic magnetic resonance imaging (µMRI) and polarized light microscopy (PLM) are used to characterize the structural variations at different anatomical locations of femoral cartilage in young rabbits (12-14 weeks old).DesignFour intact knees were imaged by µMRI at 86 µm resolution. Three small cartilage-bone specimens were harvested from each of 2 femoral medial condyles and imaged by quantitative µMRI (T2 anisotropy) at 9.75 µm resolution (N = 6). These specimens, as well as the other 2 intact femoral condyles, were used for histology and imaged by quantitative PLM (retardation and angle) at 0.25 µm to 4 µm resolutions.ResultsQuantitative MRI relaxation data and PLM fibril data revealed collaboratively distinct topographical variations in both cartilage thickness and its collagen organization in the juvenile joint. Cartilage characteristics from the central location have a 3-zone arcade-like fibril structure and a distinct magic angle effect, commonly seen in mature articular cartilage, while cartilage at the anterior location lacks these characteristics. Overall, the lowest retardation values and isotropic T2 values have been found in the distal femur (trochlear ridge), with predominant parallel fibers with respect to the articular surface. Central cartilage is the thickest (~550 µm), approximately twice as thick as the anterior and posterior locations.ConclusionDistinctly different characteristics of tissue properties were found in cartilage at different topographical locations on femoral condyle in rabbits. Knowledge of location-specific structural differences in the collagen network over the joint surface can improve the understanding of local mechanobiology and provide insights to tissue engineering and degradation repairs.

In silico and in vivo studies of the effect of surface curvature on the osteoconduction of porous scaffolds.

Recent evidence shows that the curvature of porous scaffold plays a significant role in guiding tissue regeneration. However, the underlying mechanism remains controversial to date. In this study, we developed an in silico model to simulate the effect of surface curvature on the osteoconduction of scaffold implants, which comprises the primary aspects of bone regeneration. Selective laser melting was used to manufacture a titanium scaffold with channels representative of different strut curvatures for in vivo assessment. The titanium scaffold was implanted in the femur condyles of rabbits to validate the mathematical model. Simulation results suggest that the curvature affected the distribution of growth factors and subsequently induced the migration of osteoblast lineage cells and bone deposition to the locations with higher curvature. The predictions of the mathematical model are in good agreement with the in vivo assessment results, in which newly formed bone first appeared adjacent to the vertices of the major axes in elliptical channels. The mechanism of curvature-guided osteoconduction may provide a guide for the design optimization of scaffold implants to achieve enhanced bone ingrowth.

ПРОЛОНГАЦИЯ ОЧИЩЕНИЯ ПОВРЕЖДЕННЫХ ТКАНЕЙ ОТ ДЕТРИТА В УСЛОВИЯХ ПРИМЕНЕНИЯ ЭКЗОСОМ МУЛЬТИПОТЕНТНЫХ СТРОМАЛЬНЫХ КЛЕТОК

Objective. To study the effect of exosomes of multipotent mesenchymal stromal cells (EMSCs) on soft tissues damaged during implantation of a metal screw into the bone. Methods. A defect (2 mm in diameter and 4 mm in depth) was created in the tibial proximal condyles of outbred rabbits. Metal screws were implanted into the defect by preliminary injection of saline (control, n=9 animals) or 19.2 μg of EMSCs per limb (experiment, n=10 rabbits). After 3, 7 and 10 days following the operation, the animals were taken out from the experiment; histological sections of soft tissues from the condyle surface, stained by hematoxylin and eosin were studied using light microscopy. Results. The use of water cooling in the process of introducing the metal implant into the tibial proximal condyle does not lead to complete removal of small bone fragments, which are subsequently either eliminated outward with wound discharge, or are destroyed and are subjected to lysis by macrophages. As a result of the EMSC effect on soft tissues near the site of damage, the activity of the postoperative inflammation reduces, leads to a slowdown in the resorption of hemorrhages, the elimination of fibrin clots, detritus and small bone fragments. Even on the 10&lt;sup&gt;th&lt;/sup&gt; day after using EMSCs in the postoperative wound a structureless detritus with a small number of infiltrating cells was present, as well as a significant number of multinucleated macrophages with fused cytoplasm, non-viable lysed striated muscle symplasts and bone fragments with a low degree of degradation. Conclusion. Suppression of inflammation by EMSCs delays the clearance of the postoperative wound, promotes the prolongation of the repair process and the attachment of the granulomatous component to the inflammation. The using EMSCs in the process of intraosseous implantation may be recommended only to control the activity of the inflammatory process and only after maximum preliminary cleansing of the postoperative wound from detritus, including non-viable muscle tissue and bone fragments. What this paper adds It has been firstly shown that after the experimental use of exosome of multipotent mesenchymal stromal cells to influence the regeneration of surgical trauma of soft tissues after intraosseous implantation, it is possible to reduce the activity of the inflammatory reaction, which leads to a slowdown of resorption of hemorrhages, elimination of fibrin clots, detritus and small bone fragments and, accordingly, prolongation of cleansing damaged tissue and delayed repair.

Insight Into Osseointegration of Nanohydroxyapatite/Polyamide 66 Based on the Radiolucent Gap: Comparison With Polyether-Ether-Ketone

Spinal fusion cages have been used in spinal fusion surgery for over 20 years. Polyether-ether-ketone (PEEK) cages are one of the most widely used materials. However, an increasing number of clinical and preclinical studies have shown that as a bioinert material the PEEK cage causes implant failure owing to limited osseointegration. The most common complication is a radiolucent zone at the bone-implant interface. Nanohydroxyapatite/polyamide 66 (n-HA/PA66) is a bioactive composite with sufficient load-bearing properties and good osseointegration abilities. However, in the early stage after surgery, a radiolucent gap can also be observed at the margin of the bone-implant interface. To better assess osseointegration performance as a fusion cage and compare the radiolucent gaps between the two materials, PEEK and n-HA/PA66, implants were prepared and implanted into the femoral condyles of adult New Zealand white rabbits to create a line-to-line bone-implant interface model. The interfaces were systematically investigated using X-ray radiography, histological analysis, scanning electron microscopy (SEM), elemental mapping analysis, micro-computed tomography evaluation, and push-out tests at 4, 8, 12, 24, and 52 weeks. Analysis of X-ray films and histological sections indicated a radiolucent gap around the margin of n-HA/PA66 in the early weeks after implantation (4–8 weeks). The gap narrowed and decreased gradually at 24–52 weeks. Histological analysis and SEM suggested that the formed bone could integrate and adhere in some regions of the implant surface. In addition, a better bone-like apatite layer was formed between the bone and the n-HA/PA66 implant interface than with the PEEK implant. Push-out tests conducted at 24 and 52 weeks to evaluate integrated strength showed that the n-HA/PA66 implants have better bonding strength and sufficient stability, whereas PEEK implants possess poor integrated strength. Therefore, the n-HA/PA66 composite exhibits good osseointegration properties and an improved integrated bone-implant interface.

Microalloying Design of Biodegradable Mg-2Zn-0.05Ca Promises Improved Bone-Implant Applications.

Mg and its alloys have been comprehensively studied and show huge potential for clinical orthopedic applications. However, balancing the mechanical strength and corrosion resistance of alloys is still a challenge. In light of this, micro-level contents of Zn and Ca were added to pure Mg to fabricate a Mg-2Zn-0.05Ca microalloy to expectedly enhance the mechanical strength and concurrently improve the corrosion resistance. The characteristics of the rolled Mg-2Zn-0.05Ca microalloy were explored using optical microscopy, X-ray diffraction, and tensile tests. The corrosion behavior and mechanical strength loss were explored using electrochemical and immersion tests. The effects of the microalloy extract on the proliferation, adhesion, and osteogenic differentiation of MC3T3-E1 cells were systematically studied. Moreover, implantations were done in femoral condyles of rabbits to study the degradation properties, osteogenic effect, mechanical strength loss, and biosafety of the microalloy. The ultimate tensile strength and yield strength of the rolled microalloy were found to be significantly elevated to 257 ± 2.74 and 237.6 ± 8.29 MPa, respectively. The microalloy showed a stable and gradual strength loss during degradation, both in vivo and in vitro. Concurrently, the microalloy exhibited improved corrosion resistance ability and especially, in vivo, the rolled microalloy exhibited a comparable degradation rate to that of rolled pure Mg within the initial 12 weeks of implantation. Additionally, the microalloy promoted osteogenesis, both in vitro and in vivo, and no short- and long-term toxicities of the microalloy were observed in rabbits. This study suggested that the rolled Mg-2Zn-0.05Ca microalloy effectively balanced the mechanical strength and corrosion resistance and showed potential application as bone implants.

Osseointegration of additive manufacturing Ti–6Al–4V and Co–Cr–Mo alloys, with and without surface functionalization with hydroxyapatite and type I collagen

The introduction of additive manufacturing (AM) technologies has profoundly revolutionized the implant manufacturing industry, with a particularly significant impact on the field of orthopedics. Electron Beam Melting (EBM) and Direct Metal Laser Sintering (DMLS) represents AM fabrication techniques with a pivotal role in the realization of complex and innovative structure starting from virtual 3D model data. In this study, Ti–6Al–4V and Co–Cr–Mo materials, developed by EBM (Ti-POR) and DMLS (Co-POR) techniques, respectively, with hydroxyapatite (Ti-POR + HA; Co-POR + HA) and type I collagen (Ti-POR-COLL; Co-POR-COLL) coatings, were implanted into lateral femoral condyles of rabbits. Osseointegration process was investigated by histological, histomorphometrical and microhardness evaluations at 4 and 12 weeks after implantation. Both Ti–6Al–4V and Co–Cr–Mo implants, with or without HA and COLL coatings, demonstrated good biocompatibility. As expected, HA coating hastened bone-to-implant contact (BIC) process, while collagen did not significantly improved the osseointegration process in comparison to controls. Regarding newly trabecular bone formation (B.Ar/T.Ar), Co-POR presented the highest values, significantly different from those of Co-POR-COLL. Over time, an increase of BIC parameter and a decrease of B.Ar/T.Ar were detected. Higher mineral apposition rate was observed for Ti-POR and Co-POR in comparison to Ti-POR-COLL and Co-POR-COLL, respectively, at 12 weeks. The same behavior was found for bone formation rate between Co-POR and Co-POR-COLL at 12 weeks. In conclusion, the AM materials guarantee a good osseointegration and provide a suitable environment for bone regeneration with the peculiarity of allowing personalized and patient-specific needs customization to further improve the long-term clinical outcomes.

Assessment of the in vivo biofunctionality of a biomimetic hybrid scaffold for osteochondral tissue regeneration.

Chondral and osteochondral lesions represent one of the most challenging problems in the orthopedic field, as these types of injuries lead to disability and worsened quality of life for patients and have an economic impact on the healthcare system. The aim of this in vivo study was to develop a new tissue engineering approach through a hybrid scaffold for osteochondral tissue regeneration made of porous polyurethane foam (PU) coated under vacuum with calcium phosphates (PU/VAC). Scaffold characterization showed a highly porous and interconnected structure. Human amniotic mesenchymal stromal cells (hAMSCs) were loaded into scaffolds using pectin (PECT) as a carrier. Osteochondral defects in medial femoral condyles of rabbits were created and randomly allocated in one of the following groups: plain scaffold (PU/VAC), scaffold with hAMSCs injected in the implant site (PU/VAC/hAMSC), scaffold with hAMSCs loaded in pectin (PU/VAC/PECT/hAMSC), and no treated defects (untreated). The therapeutic efficacy was assessed by macroscopic, histological, histomorphometric, microtomographic, and ultrastructural analyses at 3, 6, 12, and 24 weeks. Histological results showed that the scaffold was permissive to tissue growth and penetration, an immature osteocartilaginous tissue was observed at early experimental times, with a more accentuated bone regeneration in comparison with the cartilage layer in the absence of any inflammatory reaction.

Bone Morphogenetic Protein-2-Activated 3D-Printed Polylactic Acid Scaffolds to Promote Bone Regrowth and Repair.

Uneven distribution of pores, lack of connection between holes, low reproducibility, insufficient mechanical strength, and incomplete volatility of organic solvents are some problems associated with traditional tissue engineering methods for bone defect repair. These characteristics reduce the quality and stability of products. This study uses 3D printing (3DP) to fabricate a biocompatible poly(lactic) acid-based scaffold for repairing bone tissue. Hence, three different types of scaffolds are assessed: a freeze-dried polylactic acid (PLA) scaffold constructed using the traditional freeze-extraction method; a 3D-PLA scaffold produced through the 3DP technique; and a 3D-PLA-bone morphogenetic protein-2 (BMP-2) scaffold that is prepared using 3DP technology, with the addition of BMP-2. To enhance biological activity, polydopamine (pDA) is used to graft BMP-2 on the surface of the 3D-PLA-BMP-2 scaffold. Then, the scaffolds are implanted into the bilateral femoral condyles of rabbits, and their ability to repair the bone tissue defects is tested. The results of the experiments reveal that the 3DP scaffolds are more biocompatible than the ones produced through the traditional manufacturing methods because they enhance cell adhesion and differentiation after pDA modification and BMP-2 fixation. In the future, the 3DP products may be applied for the repair of larger bone defects in the clinical setting.