Platelet-Rich Plasma-Loaded Poly(d,l-lactide)-Poly(ethylene glycol)-Poly(d,l-lactide) Hydrogel Dressing Promotes Full-Thickness Skin Wound Healing in a Rodent Model.

Traditional therapeutic methods for skin wounds have many disadvantages, and new wound dressings that can facilitate the healing process are thus urgently needed. Platelet-rich plasma (PRP) contains multiple growth factors (GFs) and shows a significant capacity to heal soft tissue wounds. However, these GFs have a short half-life and deactivate rapidly; we therefore need a sustained delivery system to overcome this shortcoming. In this study, poly(d,l-lactide)-poly(ethylene glycol)-poly(d,l-lactide) (PDLLA-PEG-PDLLA: PLEL) hydrogel was successfully created as delivery vehicle for PRP GFs and was evaluated systematically. PLEL hydrogel was injectable at room temperature and exhibited a smart thermosensitive in situ gel-formation behavior at body temperature. In vitro cell culture showed PRP-loaded PLEL hydrogel (PRP/PLEL) had little cytotoxicity, and promoted EaHy926 proliferation, migration and tube formation; the factor release assay additionally indicated that PLEL realized the controlled release of PRP GFs for as long as 14 days. When employed to treat rodents' full-thickness skin defects, PRP/PLEL showed a significantly better ability to raise the number of both newly formed and mature blood vessels compared to the control, PLEL and PRP groups. Furthermore, the PRP/PLEL-treated group displayed faster wound closure, better reepithelialization and collagen formation. Taken together, PRP/PLEL provides a promising strategy for promoting angiogenesis and skin wound healing, which extends the potential of this dressing for clinical application.

Adipose-derived stem cell-based optimization strategies for musculoskeletal regeneration: recent advances and perspectives.

Musculoskeletal disorders are the leading causes of physical disabilities worldwide. The poor self-repair capacity of musculoskeletal tissues and the absence of effective therapies have driven the development of novel bioengineering-based therapeutic approaches. Adipose-derived stem cell (ADSC)-based therapies are being explored as new regenerative strategies for the repair and regeneration of bone, cartilage, and tendon owing to the accessibility, multipotency, and active paracrine activity of ADSCs. In this review, recent advances in ADSCs and their optimization strategies, including ADSC-derived exosomes (ADSC-Exos), biomaterials, and genetic modifications, are summarized. Furthermore, the preclinical and clinical applications of ADSCs and ADSC-Exos, either alone or in combination with growth factors or biomaterials or in genetically modified forms, for bone, cartilage, and tendon regeneration are reviewed. ADSC-based optimization strategies hold promise for the management of multiple types of musculoskeletal injuries. The timely summary and highlights provided here could offer guidance for further investigations to accelerate the development and clinical application of ADSC-based therapies in musculoskeletal regeneration.

Three-Dimensional Bioprinting of a Structure-, Composition-, and Mechanics-Graded Biomimetic Scaffold Coated with Specific Decellularized Extracellular Matrix to Improve the Tendon-to-Bone Healing.

Healing of a damaged tendon-to-bone enthesis occurs through the formation of fibrovascular scar tissue with greatly compromised histological and biomechanical properties instead of the regeneration of a new enthesis due to the lack of graded tissue-engineering zones in the interface during the healing process. In the present study, a structure-, composition-, and mechanics-graded biomimetic scaffold (GBS) coated with specific decellularized extracellular matrix (dECM) (GBS-E) aimed to enhance its cellular differentiation inducibilities was fabricated using a three-dimensional (3-D) bioprinting technique. In vitro cellular differentiation studies showed that from the tendon-engineering zone to the bone-engineering zone in the GBS, the tenogenic differentiation inducibility decreased in correspondence with an increase in the osteogenic differentiation inducibility. The chondrogenic differentiation inducibility peaked in the middle, which was in consistent with the graded cellular phenotypes observed in a native tendon-to-bone enthesis, while specific dECM coating from the tendon-engineering zone to the bone-engineering zone (tendon-, cartilage-, and bone-derived dECM, respectively) further enhanced its cellular differentiation inducibilities (GBS-E). In a rabbit rotator cuff tear model, histological analysis showed that the GBS-E group exhibited well-graded tendon-to-bone differentiated properties in the repaired interface that was similar to a native tendon-to-bone enthesis at 16 weeks. Moreover, the biomechanical properties in the GBS-E group were also significantly higher than those in other groups at 16 weeks. Therefore, our findings suggested a promising tissue-engineering strategy for the regeneration of a complex enthesis using a three-dimensional bioprinting technique.

Anti-Inflammatory and Prochondrogenic In Situ-Formed Injectable Hydrogel Crosslinked by Strontium-Doped Bioglass for Cartilage Regeneration.

Directed differentiation of bone marrow mesenchymal stem cells (BMSCs) toward chondrogenesis plays a predominant role in cartilage repair. However, the uncontrolled inflammatory response to implants is found to impair the stability of scaffolds and the cartilage regeneration outcome. Herein, we fabricated an injectable hydrogel crosslinked by strontium-doped bioglass (SrBG) to modulate both human BMSC (hBMSC) differentiation and the inflammatory response. The results revealed that the introduction of Sr ions could simultaneously enhance the proliferation of hBMSCs, upregulate cartilage-specific gene expression, and improve the secretion of glycosaminoglycan. Moreover, after cultured with SA/SrBG extracts in vitro, a majority of macrophages were polarized toward the M2 phenotype and subsequently facilitated the chondrogenic differentiation of hBMSCs. Furthermore, after the composite hydrogel was injected into a cartilage defect model, neonatal cartilage-like tissues with a smooth surface and tight integration with original tissues could be found. This study suggests that the synergistic strategy based on an enhanced differentiation ability and a regulated inflammatory response is promising and may lead the way to new anti-inflammatory biomaterials.

Multilayer Injectable Hydrogel System Sequentially Delivers Bioactive Substances for Each Wound Healing Stage.

Wound healing is a dynamic and complex process that contains several sequential phases. However, most of the current drug delivery systems were designed to treat only one certain phase of wound repair, ignoring the fact that every stage plays critical roles in the wound healing process and those critical stages coordinately work to ensure optimal tissue regeneration. Therefore, a delivery system that can precisely meet the requirements of each wound healing stage is desired to enhance tissue regeneration. In this study, an injectable sodium alginate/bioglass (SA/BG) composite hydrogel was used to carry SA microparticles containing a conditioned medium (CM) of cells (SACM). Inside the SACM microparticles, poly(lactic-co-glycolic acid) (PLGA) microspheres containing pirfenidone (PFD) were encapsulated (PLGAPFD). This multilayer injectable hydrogel system (SA/BG-SACM-PLGAPFD) was designed to sequentially deliver bioactive molecules for meeting the bioactivity requirement and timeline of each wound healing stage. First, SA/BG hydrogels could rapidly release BG ionic products in the first 1-3 days to regulate the inflammatory response of the host and initiate the subsequent tissue regeneration. Then, SACM hydrogel microparticles could release CM of RAW 264.7 cells stimulated with BG ionic products in 2-7 days to facilitate the formation of the vascularized granulation tissue. Finally, PLGAPFD microspheres released PFD in 8-20 days to prevent the fibrosis and scar formation in the regenerated skin. Thus, this SA/BG-SACM-PLGAPFD delivery system could restrain host inflammation, accelerate wound healing, and inhibit the fibrosis formation in a diabetic mouse skin damage model, enhancing skin regeneration. As the bioactive components in each layer of the system can be adjusted according to the requirements of different tissue regeneration, this three-layered injectable biomaterial system has a wide application potential in the regenerative medicine field.

Pro-chondrogenic and immunomodulatory melatonin-loaded electrospun membranes for tendon-to-bone healing.

Reconstructing the native structure of the tendon-to-bone insertion site (enthesis) in rotator cuff repair has always been a great challenge for orthopedic surgeons. Difficulty arises mainly due to the limited enthesis regenerative capability and severe inflammatory cell infiltration, which result in fibrovascular scar formation instead of native cartilage-like enthesis. Therefore, tissue engineering scaffolds with pro-chondrogenic and immunomodulatory capabilities may offer a new strategy for native enthesis regeneration. In this study, melatonin-loaded aligned polycaprolactone (PCL) electrospun fibrous membranes were fabricated. The sustained release of melatonin from this membrane significantly promoted the chondrogenic differentiation of human bone marrow-derived mesenchymal stem cells (hBMSCs) in a long-term chondroid pellet model. After the membranes were implanted in a rat acute rotator cuff tear model, melatonin-loaded PCL membranes inhibited macrophage infiltration in the tendon-to-bone interface at the early healing phase, increasing chondroid zone formation, promoting collagen maturation, decreasing fibrovascular tissue formation and eventually improving the biomechanical strength of the regenerated enthesis. Taken together, melatonin-loaded PCL membranes possess great clinical application potential for tendon-to-bone healing.

Enhancement of tendon­bone healing following rotator cuff repair using hydroxyapatite with TGFß1.

The formation of fibrocartilage at the healing site following a rotator cuff tear repair is a major problem in the field of tendon­bone healing. The present study aimed to enhance the healing of the tendon­bone interface following rotator­cuff tear repair by the interposition of hydroxyapatite (HA) encapsulated with transforming growth factor ß1 (TGFß1). Using an acute rotator cuff repair model, rats were divided into three groups: i) Repair only (control); ii) HA group; and iii) HA­TGFß1 group. Animals were sacrificed at 2, 4 and 8 weeks following surgery. Micro­computed tomography (CT), histomorphometric analyses and biomechanical tests were used to evaluate the supraspinatus tendon­bone complex. The micro­CT images revealed notable novel bone formation in the groups treated with HA­TGFß1. The histomorphometric analyses demonstrated improved fibrocartilage formation and collagen organization at the tendon­bone interface. The HA­TGFß1 combination significantly improved the area of fibrocartilage, particularly at early time points (2 and 4 weeks). There was a significantly greater load­to­failure force achieved in the HA and HA­TGFß1 groups compared with the control group at 4 and 8 weeks. Augmentation of the ceramic powder with HA­TGFß1 at the tendon­bone interface was demonstrated to strengthen the healing entheses, increase bone and fibrocartilage formation and improve collagen organization compared with surgical repair alone. Local application of HA­TGFß1 demonstrates potential in improving tendon­bone healing following rotator cuff repair.

Evaluation of zinc-doped mesoporous hydroxyapatite microspheres for the construction of a novel biomimetic scaffold optimized for bone augmentation.

Biomaterials with high osteogenic activity are desirable for sufficient healing of bone defects resulting from trauma, tumor, infection, and congenital abnormalities. Synthetic materials mimicking the structure and composition of human trabecular bone are of considerable potential in bone augmentation. In the present study, a zinc (Zn)-doped mesoporous hydroxyapatite microspheres (Zn-MHMs)/collagen scaffold (Zn-MHMs/Coll) was developed through a lyophilization fabrication process and designed to mimic the trabecular bone. The Zn-MHMs were synthesized through a microwave-hydrothermal method by using creatine phosphate as an organic phosphorus source. Zn-MHMs that consist of hydroxyapatite nanosheets showed relatively uniform spherical morphology, mesoporous hollow structure, high specific surface area, and homogeneous Zn distribution. They were additionally investigated as a drug nanocarrier, which was efficient in drug delivery and presented a pH-responsive drug release behavior. Furthermore, they were incorporated into the collagen matrix to construct a biomimetic scaffold optimized for bone tissue regeneration. The Zn-MHMs/Coll scaffolds showed an interconnected pore structure in the range of 100-300 µm and a sustained release of Zn ions. More importantly, the Zn-MHMs/Coll scaffolds could enhance the osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells. Finally, the bone defect repair results of critical-sized femoral condyle defect rat model demonstrated that the Zn-MHMs/Coll scaffolds could enhance bone regeneration compared with the Coll or MHMs/Coll scaffolds. The results suggest that the biomimetic Zn-MHMs/Coll scaffolds may be of enormous potential in bone repair and regeneration.

In vitro and in vivo evaluation of MgF2 coated AZ31 magnesium alloy porous scaffolds for bone regeneration.

Porous magnesium scaffolds are attracting increasing attention because of their degradability and good mechanical property. In this work, a porous and degradable AZ31 magnesium alloy scaffold was fabricated using laser perforation technique. To enhance the corrosion resistance and cytocompatibility of the AZ31 scaffolds, a fluoride treatment was used to acquire the MgF2 coating. Enhanced corrosion resistance was confirmed by immersion and electrochemical tests. Due to the protection provided by the MgF2 coating, the magnesium release and pH increase resulting from the degradation of the FAZ31 scaffolds were controllable. Moreover, in vitro studies revealed that the MgF2 coated AZ31 (FAZ31) scaffolds enhanced the proliferation and attachment of rat bone marrow stromal cells (rBMSCs) compared with the AZ31 scaffolds. In addition, our present data indicated that the extract of the FAZ31 scaffold could enhance the osteogenic differentiation of rBMSCs. To compare the in vivo bone regenerative capacity of the AZ31 and FAZ31 scaffolds, a rabbit femoral condyle defect model was used. Micro-computed tomography (micro-CT) and histological examination were performed to evaluate the degradation of the scaffolds and bone volume changes. In addition to the enhanced the corrosion resistance, the FAZ31 scaffolds were more biocompatible and induced significantly more new bone formation in vivo. Conversely, bone resorption was observed from the AZ31 scaffolds. These promising results suggest potential clinical applications of the fluoride pretreated AZ31 scaffold for bone tissue repair and regeneration.

Cellular response of chondrocytes to magnesium alloys for orthopedic applications.

In the present study, the effects of Mg-Nd-Zn-Zr (JDBM), brushite (CaHPO4·2H2O)-coated JDBM (C-JDBM), AZ31, WE43, pure magnesium (Mg) and Ti alloy (TC4) on rabbit chondrocytes were investigated in vitro. Adhesion experiments revealed the satisfactory morphology of chondrocytes on the surface of all samples. An indirect cytotoxicity test using MTT assay revealed that C­JDBM and TC4 exhibited results similar to those of the negative control, better than those obtained with JDBM, AZ31, WE43 and pure Mg (p<0.05). There were no statistically significant differences observed between the JDBM, AZ31, WE43 and pure Mg group (p>0.05). The results of indirect cell cytotoxicity and proliferation assays, as well as those of apoptosis assay, glycosaminoglycan (GAG) quantification, assessment of collagen Ⅱ (Col Ⅱ) levels and RT-qPCR revealed a similar a trend as was observed with MTT assay. These findings suggested that the JDBM alloy was highly biocompatible with chondrocytes in vitro, yielding results similar to those of AZ31, WE43 and pure Mg. Furthermore, CaHPO4·2H2O coating significantly improved the biocompatibility of this alloy.

Enhanced biocorrosion resistance and biocompatibility of degradable Mg-Nd-Zn-Zr alloy by brushite coating.

To further improve the corrosion resistance and biocompatibility of Mg-Nd-Zn-Zr alloy (JDBM), a biodegradable calcium phosphate coating (Ca-P coating) with high bonding strength was developed using a novel chemical deposition method. The main composition of the Ca-P coating was brushite (CaHPO4·2H2O). The bonding strength between the coating and the JDBM substrate was measured to be over 10 MPa, and the thickness of the coating layer was about 10-30 µm. The in vitro corrosion tests indicated that the Ca-P treatment improved the corrosion resistance of JDBM alloy in Hank's solution. Ca-P treatment significantly reduced the hemolysis rate of JDBM alloy from 48% to 0.68%, and induced no toxicity to MC3T3-E1 cells. The in vivo implantation experiment in New Zealand's rabbit tibia showed that the degradation rate was reduced obviously by the Ca-P treatment and less gas was produced from Ca-P treated JDBM bone plates and screws in early stage of the implantation, and at least 10weeks degradation time can be prolonged by the present coating techniques. Both Ca-P treated and untreated JDBM Mg alloy induced bone growth. The primary results indicate that the present Ca-P treatment is a promising technique for the degradable Mg-based biomaterials for orthopedic applications.

Medial patellofemoral ligament reconstruction using semitendinosus tendons: polyester suture augmentation versus nonaugmentation.

BACKGROUND: The comparative clinical outcome of medial patellofemoral ligament reconstruction (MPFLR) using semitendinosus tendons with and without polyester suture augmentation for recurrent patellar instability is unknown. HYPOTHESIS: Medial patellofemoral ligament reconstruction with polyester suture augmentation will yield better results than MPFLR without augmentation for recurrent patellar instability in adults. STUDY DESIGN: Randomized controlled trial; Level of evidence, 2. METHODS: One hundred patients with recurrent patellar instability receiving MPFLR using semitendinosus tendons were randomly divided into 2 groups either with or without polyester suture augmentation. Tibial tubercle transfer was performed in most cases. Follow-ups were performed at 12, 24, and 60 months postoperatively, and computed tomography (CT) was performed immediately after the operation and at follow-up. The passive patellar glide test was performed before surgery, immediately after MPFLR during the operation, and at each follow-up point. The degree of knee function was evaluated preoperatively and at 2 and 5 years postoperatively using the International Knee Documentation Committee, Lysholm, and Kujala rating scales. Redislocation or multiple episodes of patellar instability were considered failures. RESULTS: Forty-two patients in the augmentation group and 43 patients in the nonaugmentation group were followed for 5 years and received complete serial CT examinations and functional evaluations. The correction of the static patellar position deteriorated over time in the nonaugmentation group but not in the augmentation group. The results of the passive patellar glide test indicated stable patellae in all patients immediately after MPFLR and more stable patellae in the augmentation group at each follow-up point. Functional evaluations at 2 and 5 years revealed statistically significant superior results in the augmentation group. Finally, no patient in the augmentation group and 2 (4.7%) in the nonaugmentation group experienced episodes of redislocation, and 1 patient (2.4%) in the augmentation group and 8 (18.6%) in the nonaugmentation group experienced multiple episodes of patellar instability, resulting in failure rates of 2.4% and 23.3% in the augmentation group and nonaugmentation group, respectively (P = .004). CONCLUSION: Medial patellofemoral ligament reconstruction with polyester suture augmentation results in better static patellar position, dynamic stability, and functional outcome than without augmentation in the treatment of recurrent patellar dislocation in adults.

In vivo degradation behavior and biocompatibility of Mg-Nd-Zn-Zr alloy at early stage.

The neotype magnesium alloy, Mg-Nd-Zn-Zr (NZK) alloy, was implanted into the rabbit femur to investigate its in vivo degradation behavior and biocompatibility. Seventy-two New Zealand white rabbits were randomly divided into the NZK alloy group, titanium alloy group and sham-operated group. Then NZK alloy rods were embedded in the rabbit femur in the NZK alloy group, titanium alloy rods were embedded in the titanium alloy group, and only bone tunnel was established in the sham-operated group. Prior to surgery and at 1, 7, 14, 28 and 56 days after operation, the serum alanine transaminase, creatinine, creatine kinase and magnesium ion concentration were examined in each group. An X-ray of the implanted region was taken at 7, 14, 28 and 56 days after implantation. The pathological changes in heart, liver, kidney and bone from the implant region were examined at 28 and 56 days postoperatively. The degradation behavior of the NZK alloy was observed using scanning electron microscope with an energy dispersive spectroscopy system. There were no significant differences in serum alanine transaminase, creatinine, creatine kinase and magnesium ion concentrations among each group at the same time point (P>0.05). The histology of heart, liver, kidney and bone from implant region was altered. The results demonstrate that the NZK alloy implanted into the rabbit femur could be absorbed gradually, and that the NZK alloy has excellent biocompatibility in vivo.

Biocompatibility of magnesium-zinc alloy in biodegradable orthopedic implants.

In this study, magnesium-zinc (Mg-Zn) alloy was investigated as a biodegradable orthopedic implant. MC3T3-E1 cell attachment, mineralization and osteogenic-specific mRNA expression were assessed for as measurements of the in vitro biocompatibility of Mg-Zn alloy. In vivo degradation of the Mg-Zn alloy and the accompanying new bone formation in the femoral marrow cavity were analyzed by scanning electron microscopy and histomorphological analysis. Results showed that MC3T3-E1 cells cultured on Mg-Zn alloy samples manifested better attachment and mineralization ability, as well as improved mRNA expression of collagen 1 α 1 (COL1α1) and osteocalcin (OC), compared with cells seeded on poly-L-lactic acid (PLLA) samples. In vivo experiments demonstrated that, compared with PLLA materials, the Mg-Zn alloy not only degraded faster, but was accompanied by considerable new bone formation around the samples. Our data indicate that Mg-Zn alloy has excellent biocompatibility for application as degradable bone implants.

Research on an Mg-Zn alloy as a degradable biomaterial.

In this study a binary Mg-Zn magnesium alloy was researched as a degradable biomedical material. An Mg-Zn alloy fabricated with high-purity raw materials and using a clean melting process had very low levels of impurities. After solid solution treatment and hot working the grain size of the Mg-Zn alloy was finer and a uniform single phase was gained. The mechanical properties of this Mg-Zn alloy were suitable for implant applications, i.e. the tensile strength and elongation achieved were approximately 279.5MPa and 18.8%, respectively. The results of in vitro degradation experiments including electrochemical measurements and immersion tests revealed that the zinc could elevate the corrosion potential of Mg in simulated body fluid (SBF) and reduce the degradation rate. The corrosion products on the surface of Mg-Zn were hydroxyapatite (HA) and other Mg/Ca phosphates in SBF. In addition, the influence caused by in vitro degradation on mechanical properties was studied, and the results showed that the bending strength of Mg-Zn alloy dropped sharply in the earlier stage of degradation, while smoothly during the later period. The in vitro cytotoxicity of Mg-Zn was examined. The result 0-1 grade revealed that the Mg-Zn alloy was harmless to L-929 cells. For in vivo experiments, Mg-Zn rods were implanted into the femoral shaft of rabbits. The radiographs illustrated that the magnesium alloy could be gradually absorbed in vivo at about 2.32mm/yr degradation rate obtained by weight loss method. Hematoxylin and eosin (HE) stained section around Mg-Zn rods suggested that there were newly formed bone surrounding the implant. HE stained tissue (containing heart, liver, kidney and spleen tissues) and the biochemical measurements, including serum magnesium, serum creatinine (CREA), blood urea nitrogen (BUN), glutamic-pyruvic transaminase (GPT) and creatine kinase (CK) proved that the in vivo degradation of Mg-Zn did not harm the important organs. Moreover, no adverse effects of hydrogen generated by degradation had been observed and also no negative effects caused by the release of zinc were detected. These results suggested that the novel Mg-Zn binary alloy had good biocompatibility in vivo.