Evaluation of natural protein-based nanofiber composite photocrosslinking hydrogel for skin wound regeneration.

Protein based photocrosslinking hydrogels with nanofiber dispersions were reported to be an effective wound dressing. In this study, two kinds of protein (gelatin and decellularized dermal matrix) were modified to obtain GelMA and ddECMMA, respectively. Poly(ε-caprolactone) nanofiber dispersions (PCLPBA) and thioglycolic acid-modified chitosan (TCS) were added into GelMA solution and ddECMMA solution, respectively. After photocrosslinking, four kinds of hydrogel (GelMA, GTP4, DP and DTP4) were fabricated. The hydrogels showed excellent physico-chemical property, biocompatibility and negligible cytotoxicity. When applied on the full-thickness cutaneous deficiency of SD rats, hydrogel treated groups exhibited an enhanced wound healing effect than Blank group. Besides, the histological staining of H&E and Masson's showed that hydrogels groups with PCLPBA and TCS (GTP4 and DTP4) improved wound healing. Furthermore, GTP4 group performed better healing effect than other groups, which had great potential in skin wound regeneration.

The role of vitamin D on rotator cuff tear with osteoporosis.

Backgrounds: Osteoporosis (OP) is an important risk factor for rotator cuff tears (RCTs). However, the relationship and mechanism between rotator cuff injury and osteoporosis are unclear. Therefore, to investigate association between rotator cuff injury and osteoporosis, and find clinical characteristics, bone mineral density, bone metabolism markers, and nutrient levels in rotator cuff injury patients with or without osteoporosis. Methods: One hundred and four cases of patients (RCTs, n=32; RCTs-OP, n=72) who underwent rotator cuff injury and need arthroscopic rotator cuff repair between June 2021 and February 2022, along with the diagnosis of osteoporosis were identified from the dual-energy X-ray bone density screening(DXA). The outcome measure includes clinical characteristics, bone mineral density, bone metabolism markers, vitamins, and amino acids. Multivariable logistic regression analysis was applied to build a predicting model incorporating the feature selected in the least absolute shrinkage and selection operator regression model. Discrimination, calibration, and clinical usefulness of the predicting model were assessed using the C-index, calibration plot, and decision curve analysis. Internal validation was assessed using bootstrapping validation. Results: OP with RCTs has a lower level of in 25-vitD, osteocalcin (OCN), serum Ca2+, ornithine, diaminocaproic_acid but the high level of Vitamin_B12, PTH, Vitamin_D3,γ_aminobutyric_acid, Vitamin_C and Vitamin_E than RCTs patients without OP. Predictors contained in the prediction nomogram included lumber T score, femur T score, Niacin_B3, and vitamin D, reflecting the combined effect of vitamins on RCTs-related OP progression. The model has good discriminative ability with a C-index of 0.938(95% CI:-1.83-1.39) and good scaling ability. The high C-index value of 0.95 is still achievable with range validation. Analysis of decision curves showed that non-adherence is clinically useful when intervention decisions are at the 14% probability limit of non-adherence. Conclusion: This study supports the hypothesis that lumber T score, femur T score, Niacin_B3, and Vitamin D are valuable prognostic biomarkers on RCTs related OP progression. What is known about the subject: It is found that vitamin D are valuable prognostic biomarkers, reflecting the combined effect of vitamins on RCTs related OP progression. What this study adds to existing knowledge: These findings also highlight that nutrients condition such as vitamins and amino acids of patients provide a new understanding of the development of RCTs.

Arthroscopic Modified Double-Row Biceps Tenodesis versus Labral Repair for the Treatment of Isolated Type II SLAP Lesions in Non-Overhead Athletes.

OBJECTIVE: To evaluate the postoperative efficacy and the clinical outcomes of arthroscopic modified double-row biceps tenodesis versus labral repair. METHODS: A retrospective study was conducted in 56 patients with isolated type II superior labrum anterior and posterior (SLAP) lesions from March 2015 to November 2018. Thirty patients (male:female = 17:13) were treated with labral repair, and 26 patients (male:female = 15:11) were treated with modified double-row biceps tenodesis. The average age of the labral repair group and the modified double-row biceps tenodesis group were 42.8 ± 10.6 and 40.9 ± 10.2 years, respectively. Pre- and postoperative assessments with the visual analog scale (VAS), University of California Los Angeles (UCLA), and American Shoulder and Elbow Surgeons (ASES) scores were compared between the two treatment groups. Additional outcome measures included patient satisfaction, the time to return to previous activities, workers' compensation status, and postoperative complications. RESULTS: At a 2-year follow-up, the tenodesis group showed significant differences in postoperative VAS (1.5 to 1.8, respectively; p = 0.008), patient satisfaction (92.3% vs. 46.7%, p < 0.001), and recovery time to return to their previous activities (6.8 ± 1.8 vs. 8.1 ± 1.5, p = 0.007) compared to the labral repair group; however, there was no significant difference in postoperative ASES and UCLA scores between the two groups. Additionally, one patient in the tenodesis group developed persistent postoperative stiffness, which was resolved by conservative treatment. In the labral repair group, two patients presented with persistent postoperative night pain, three developed persistent postoperative stiffness, and two required a subsequent capsular release. CONCLUSIONS: Compared with the labral repair group, the arthroscopic modified double-row biceps tenodesis showed more encouraging postoperative pain reduction, earlier recovery to previous activities, and higher patient satisfaction.

Effects of preoperative serum vitamin D levels on early clinical function outcomes and the moderate-to-severe pain prevalence in postmenopausal women after primary total knee arthroplasty.

OBJECTIVE: To investigate the impact of vitamin D levels on early clinical function outcomes and the potential risk factors of moderate-to-severe pain prevalence in postmenopausal women after primary total knee arthroplasty (TKA). METHODS: From April 2017 to December 2019, 226 women were retrospectively recruited. The women were divided into two groups based on their preoperative serum 25-hydroxyvitamin D levels: (1) vitamin D-sufficient group (≥30 ng/mL); (2) vitamin D-deficient group (<30 ng/mL). The visual analog scale, Western Ontario and McMaster Arthritis Index score, and Knee Society Score were used to evaluate clinical outcomes. Risk factors for developing postoperative moderate-to-severe knee pain were studied using multivariate binary logistic regression analyses. RESULTS: There was no significant difference in preoperative clinical function assessment between the two groups. The difference in postoperative Western Ontario and McMaster Arthritis Index score between the two groups was statistically significant (15.3 ±â€Š0.7 vs 15.6 ±â€Š0.7: P = 0.02). However, the differences in postoperative visual analog scale and Knee Society Score scores between the two groups were not significant (P > 0.05). The incidence of postoperative moderate-to-severe pain was 16.4% (95% CI 11.8%-21.9%). Multivariate logistic regression analysis revealed that vitamin D deficiency, smoking, and high body mass index were potential risk factors for moderate-to-severe knee pain in postmenopausal women early after TKA (P < 0.05). CONCLUSION: Preoperative vitamin D deficiency may adversely affect early functional outcomes in postmenopausal women after TKA. In addition, vitamin D deficiency, smoking, and high body mass index were independent risk factors for moderate-to-severe knee pain after surgery.

MicroRNA­218 promotes prostaglandin E2 to inhibit osteogenic differentiation in synovial mesenchymal stem cells by targeting 15­hydroxyprostaglandin dehydrogenase [NAD(+)].

The chondrogenic differentiation of synovial mesenchymal stem cells (SMSCs) is regulated by essential transcription factors and signaling cascades. However, the precise mechanisms involved in this process remain unclear. MicroRNAs (miRs/miRNAs) are undersized non­coding RNAs responsible for the post­transcriptional regulation of gene expression, by binding to the 3'­untranslated regions (3'­UTRs) of their target mRNAs. miRNAs may constitute a promising tool to regulate SMSC differentiation and to advance the controlled differentiation of SMSCs in therapeutic applications. The aim of the present study was to examine the role of miR­218 in SMSC differentiation towards chondrocytes. The present study comparatively analyzed the expression profile of known miRNAs and specific target genes in SMSCs between early and late differentiation stages. Western blotting and reverse transcription­quantitative polymerase chain reaction analysis of gene expression demonstrated the upregulation of 15­hydroxyprostaglandin dehydrogenase [NAD(+)] (15­HPGD), prostaglandin E2 (PGE2) and rate limiting enzymes responsible for the synthesis of PGE2 precursors throughout chondrogenesis. Through correlation analysis, it was observed that there was a significant association between miR­128, 15­HPGD gene expression, 15­HPGD protein expression and microsomal prostaglandin E synthase 1. Further experiments demonstrated that miR­218 decreased PGE2 concentration by binding to the 3'­UTR of 15­HPGD. Using an immunofluorescence reporting system, it was observed that miR­218 regulated the expression of 15­HPGD during the differentiation of SMSCs into cartilage, and subsequently inhibited osteogenesis during chondrogenesis by acting on the 3'UTR of 15­HPGD. Therefore, miR­218 may be an important regulator targeting osteogenic factors and modulating cartilage formation and differentiation. The results of the present study provided a novel insight beneficial to cellular manipulation methods during cartilage regeneration, and in cartilage tissue engineering research.

[CONDITIONS OF SYNOVIAL MESENCHYMAL STEM CELLS DIFFERENTIATING INTO FIBROCARTILAGE CELLS].

OBJECTIVE: To explore the conditions of synovial derived mesenchymal stem cells (SMSCs) differentiating into the fibrocartilage cells by using the orthogonal experiment. METHODS: The synovium was harvested from 5 adult New Zealand white rabbits, and SMSCs were separated by adherence method. The flow cytometry and multi-directional differentiation method were used to identify the SMSCs. The conditions were found from the preliminary experiment and literature review. The missing test was carried out to screen the conditions and then 12 conditions were used for the orthogonal experiment, including transforming growth factor ß1 (TGF-ß1), bone morphogenic protein 2 (BMP-2), dexamethasone (DEX), proline, ascorbic acid (ASA), pyruvic acid, insulin + transferrin + selenious acid pre-mixed solution (ITS), bovin serum albumin (BSA), basic fibroblast growth factor (bFGF), intermittent hydraulic pressure (IHP), bone morphogenic protein 7 (BMP-7), and insulin-like growth factor (IGF). The L60 (212) orthogonal experiment was designed using the SPSS 18.0 with 2 level conditions and the cells were induced to differentiate on the small intestinal submucosa (SIS)-3D scaffold. The CD151+/CD44+ cells were detected with the flow cytometry and then the differentiation rate was recorded. The immumohistochemical staining, cellular morphology, toluidine blue staining, and semi-quantitative RT-PCR examination for the gene expressions of sex determining region Y (SRY)-box 9 gene (Sox9), aggrecan gene (AGN), collagen type I gene (Col I), collagen type II gene (Col II), collagen type IX gene (Col IX) were used for result confirmation. The differentiation rate was calculated as the product of CD151/CD44+ cells and cells with Col I high expression. The grow curve was detected with the DNA abundance using the PicoGreen Assay. The visual observation and the variances analysis among the variable were used to evaluate the result of the orthogonal experiment, 1 level interaction was considered. The q-test and the least significant difference (LDS) were used for the variance analysis with a type III calibration model. The test criteria (a) was 0.05. RESULTS: The cells were certified as SMSCs, the double-time of the cells was 28 hours. During the differentiation into the fibrocartilage, the volume of the SIS-3D scaffold enlarged double every 5 days. The scaffolds were positively stained by toluidine blue at 14 days. The visual observation showed that high levels of TGF-ß1 and BMP-7 were optimum for the differentiation, and BMP-7 showed the interaction with BMP-2. The conditions of DEX, ASA, ITS, transferrin, bFGF showed decreasing promotional function by degrees, and the model showed the perfect relevance. P value was 0.000 according to the variance analysis. The intercept analysis showed different independent variables brought about variant contribution; the TGF-ß1, ASA, bFGF, IGF, and BMP-7 were more remarkable, which were similar to the visual observation. CONCLUSION: In the process of the SMSCs differentiation into the fibrocartilage, the concentrations of TGF-ß1, ASA, bFGF, and IGF reasonably can improve the conversion rate of the fibrocartilage cells. The accurate conditions of the reaulatory factor should be explored further.

Strategies to minimize hypertrophy in cartilage engineering and regeneration.

Due to a blood supply shortage, articular cartilage has a limited capacity for self-healing once damaged. Articular chondrocytes, cartilage progenitor cells, embryonic stem cells, and mesenchymal stem cells are candidate cells for cartilage regeneration. Significant current attention is paid to improving chondrogenic differentiation capacity; unfortunately, the potential chondrogenic hypertrophy of differentiated cells is largely overlooked. Consequently, the engineered tissue is actually a transient cartilage rather than a permanent one. The development of hypertrophic cartilage ends with the onset of endochondral bone formation which has inferior mechanical properties. In this review, current strategies for inhibition of chondrogenic hypertrophy are comprehensively summarized; the impact of cell source options is discussed; and potential mechanisms underlying these strategies are also categorized. This paper aims to provide guidelines for the prevention of hypertrophy in the regeneration of cartilage tissue. This knowledge may also facilitate the retardation of osteophytes in the treatment of osteoarthritis.

[Effect of transforming growth factor beta3, bone morphogenetic protein 2, and dexamethasone on chondrogenic differentiation of rabbit synovial mesenchymal stem cells].

OBJECTIVE: To study the effect of transforming growth factor beta3 (TGF-beta3), bone morphogenetic protein 2 (BMP-2), and dexamethasone (DEX) on the chondrogenic differentiation of rabbit synovial mesenchymal stem cells (SMSCs). METHODS: SMSCs were isolated from the knee joints of 5 rabbits (weighing, 1.8-2.5 kg), and were identified by morphogenetic observation, flow cytometry detection for cell surface antigen, and adipogenic and osteogenic differentiations. The SMSCs were cultured in the PELLET system for chondrogenic differentiation. The cell pellets were divided into 8 groups: TGF-beta3 was added in group A, BMP-2 in group B, DEX in group C, TGF-beta3 + BMP-2 in group C, TGF-beta3 + DEX in group E, BMP-2+ DEX in group F, and TGF-beta3 + BMP-2 + DEX in group G; group H served as control group. The diameter, weight, collagen type II (immuohistochemistry staining), proteoglycan (toluidine blue staining), and expression of cartilage related genes [real time quantitative PCR (RT-qPCR) technique] were compared to evaluate the effect of cytokines on the chondrogenic differentiation of SMSCs. Meanwhile, the DNA content of cell pellets was tested to assess the relationship between the increase weight of cell pellets and the cell proliferation. RESULTS: SMSCs were isolated from the knee joints of rabbits successfully and the findings indicated that the rabbit synovium-derived cells had characteristics of mesenchymal stem cells. The diameter, weight, collagen type II, proteoglycan, and expression of cartilage related genes of pellets in groups A-F were significantly lower than those of group G (P < 0.05). RT-qPCR detection results showed that the relative expressions of cartilage related genes (SOX-9, Aggrecan, collagen type II, collagen type X, and BMP receptor II) in group G were significantly higher than those in the other groups (P < 0.01). Meanwhile, with the increase of the volume of pellet, the DNA content reduced about 70% at 7 days, about 80% at 14 days, and about 88% at 21 days. CONCLUSION: The combination of TGF-beta3, BMP-2, and DEX can make the capacity of chondrogenesis of SMSCs maximized. The increase of the pellet volume is caused by the extracellular matrix rather than by cell proliferation.

[In vitro differentiation of synovial-derived mesenchymal stem cells infected by adenovirus vector mediated by bone morphogenetic protein 2/7 genes into fibrocartilage cells in rabbits].

OBJECTIVE: To investigate the feasibility of rabbit synovial-derived mesenchymal stem cells (SMSCs) differentiating into fibrocartilage cells by the recombinant adenovirus vector mediated by bone morphogenetic protein 2/7 (BMP-2/7) genes in vitro. METHODS: SMSCs were isolated and purified from 3-month-old New Zealand white rabbits [male or female, weighing (2.1 +/- 0.3) kg]; the morphology was observed; the cells were identified with immunocytological fluorescent staining, flow cytometry, and cell cycles. The adipogenic, osteogenic, and chondrogenic differentiations were detected. The recombinant plasmid of pAdTrack-BMP-2-internal ribosome entry site (IRES)-BMP-7 was constructed and then was used to infect SMSCs. The cell DNA content and the oncogenicity were tested to determine the safety. Then infected SMSCs were cultured in incomplete chondrogenic medium in vitro. Chondrogenic differentiation of infected SMSCs was detected by RT-PCR, immunofluorescent staining, and toluidine blue staining. RESULTS: SMSCs expressed surface markers of stem cells, and had multi-directional potential. The transfection efficiency of SMSCs infected by recombinant plasmid of pAdTrack-BMP-2-IRES-BMP-7 was about 70%. The safety results showed that infected SMSCs had normal double time, normal chromosome number, and normal DNA content and had no oncogenicity. At 21 days after cultured in incomplete chondrocyte medium, RT-PCR results showed SMSCs had increased expressions of collegan type I and collegan type II, particularly collegan type II; the expressions of RhoA and Sox-9 increased obviously. Immunofluorescent staining and toluidine blue staining showed differentiation of SMSCs into fibrocartilage cells. CONCLUSION: It is safe to use pAdTrack-BMP-2-IRES-BMP-7 for infecting SMSCs. SMSCs infected by pAdTrack-BMP-2-IRES-BMP-7 can differentiate into fibrocartilage cells spontaneously in vitro.

Reduced opioid consumption and improved early rehabilitation with local and intraarticular cocktail analgesic injection in total hip arthroplasty: a randomized controlled clinical trial.

OBJECTIVE: Postoperative pain after total hip arthroplasty (THA) is not well tolerated. We assessed postoperative pain relief and the need for opioid use after using a cocktail of local and intraarticular analgesic injection (LIA) after THA. METHODS: Eighty patients undergoing THA under spinal anesthesia were randomly assigned to receive either LIA or placebo. The LIA was composed of 5 mg morphine, 30 mg bupivacaine (15 mg/1.5 mL), 1 mL betamethasone, and 0.5 mL epinephrine (1:1,000) intraoperatively. We compared three outcomes total morphine consumption, visual analog scale (VAS) at rest and during activity, and hip flexion angle while standing. RESULTS: When compared with placebo, opioid consumption was significantly reduced in the trial group, as well as VAS at rest and during mobilization. Earlier rehabilitation and better range of motion (ROM) were achieved in the trial group. There were no significant differences in side effects or postoperative wound healing between groups. CONCLUSION: In patients undergoing THA, LIA may reduce postoperative systemic opioid use and offer better pain control and earlier rehabilitation, without observable risks.