Tung-Fu Huang

Enhancement of wound healing by human multipotent stromal cell conditioned medium: the paracrine factors and p38 MAPK activation.

Wound healing can be improved by transplanting mesenchymal stem cells (MSCs). In this study, we have demonstrated the benefits of the conditioned medium derived from human MSCs (CM-MSC) in wound healing using an excisional wound model. CM-MSC accelerated wound closure with increased reepithelialization, cell infiltration, granulation formation, and angiogenesis. Notably, CM-MSC enhanced epithelial and endothelial cell migration, suggesting the contribution of increased cell migration to wound healing enhanced by CM-MSC. Cytokine array, ELISA analysis, and quantitative RT-PCR revealed high levels of IL-6 in CM-MSC. Moreover, IL-6 added to the preconditioned medium enhanced both cell migration and wound healing, and antibodies against IL-6 blocked the increase in cell motility and wound closure by CM-MSC. The IL-6 secretory pathway of MSCs was inhibited by SB203580, an inhibitor of p38 MAPK or siRNA against p38 MAPK, suggesting IL-6 secretion by MSCs is mediated through the activation of p38 MAPK. Inactivation of p38 MAPK also reduced the expression and production of IL-8 and CXCL1 by MSCs, both of which were also demonstrated to enhance cell migration and wound closure. Thus, our data suggest MSCs promote wound healing through releasing a repertoire of paracrine factors via activation of p38 MAPK, and the CM-MSC may be applied to enhance wound healing.

Mesenchymal Stem Cells From a Hypoxic Culture Improve and Engraft Achilles Tendon Repair

Background: Bone marrow–derived mesenchymal stem cells (MSCs) from humans cultured under hypoxic conditions increase bone healing capacity. Hypothesis: Rat MSCs cultured under hypoxic conditions increase the tendon healing potential after transplantation into injured Achilles tendons. Study Design: Controlled laboratory study. Methods: Biomechanical testing, histological analysis, and bromodeoxyuridine (BrdU) labeling/collagen immunohistochemistry were performed to demonstrate that augmentation of an Achilles tendon rupture site with hypoxic MSCs increases healing capacity compared with normoxic MSCs and controls. Fifty Sprague-Dawley rats were used for the experiments, with 2 rats as the source of bone marrow MSCs. The cut Achilles tendons in the rats were equally divided into 3 groups: hypoxic MSC, normoxic MSC, and nontreated (vehicle control). The uncut tendons served as normal uncut controls. Outcome measures included mechanical testing in 24 rats, histological analysis, and BrdU labeling/collagen immunohistochemistry in another 24 rats. Results: The ultimate failure load in the hypoxic MSC group was significantly greater than that in the nontreated or normoxic MSC group at 2 weeks after incision (2.1 N/mm2 vs 1.1 N/mm2 or 1.9 N/mm2, respectively) and at 4 weeks after incision (5.5 N/mm2 vs 1.7 N/mm2 or 2.7 N/mm2, respectively). The ultimate failure load in the hypoxic MSC group at 4 weeks after incision (5.5 N/mm2) was close to but still significantly less than that of the uncut tendon (7.2 N/mm2). Histological analysis as determined by the semiquantitative Bonar histopathological grading scale revealed that the hypoxic MSC group underwent a significant improvement in Achilles tendon healing both at 2 and 4 weeks when compared with the nontreated or normoxic MSC group via statistical analysis. Immunohistochemistry further demonstrated that the hypoxic and normoxic MSC groups had stronger immunostaining for type I and type III collagen than did the nontreated group both at 2 and 4 weeks after incision. Moreover, BrdU labeling of MSCs before injection further determined the incorporation and retention of transplanted cells at the rupture site. Conclusion: Transplantation of hypoxic MSCs may be a better and more readily available treatment than normoxic MSCs for Achilles tendon ruptures. Clinical Relevance: The present study provides evidence that transplantation of hypoxic MSCs may be a promising therapy for the treatment of Achilles tendon ruptures.

Isolation of mesenchymal stem cells from shoulder rotator cuff: a potential source for muscle and tendon repair.

The self-healing potential of each tissue belongs to endogenous stem cells residing in the tissue; however, there are currently no reports mentioned for the isolation of human rotator cuff-derived mesenchymal stem cells (RC-MSCs) since. To isolate RC-MSCs, minced rotator cuff samples were first digested with enzymes and the single cell suspensions were seeded in plastic culture dishes. Twenty-four hours later, nonadherent cells were removed and the adherent cells were further cultured. The RC-MSCs had fibroblast-like morphology and were positive for the putative surface markers of MSCs, such as CD44, CD73, CD90, CD105, and CD166, and negative for the putative markers of hematopoietic cells, such as CD34, CD45, and CD133. Similar to BM-MSCs, RC-MSCs were demonstrated to have the potential to undergo osteogenic, adipogenic, and chondrogenic differentiation. Upon induction in the defined media, RC-MSCs also expressed lineage-specific genes, such as Runx 2 and osteocalcin in osteogenic induction, PPAR-γ and LPL in adipogenic differentiation, and aggrecan and Col2a1 in chondrogenic differentiation. The multipotent feature of RC-MSCs in the myogenic injury model was further strengthened by the increase in myogenic potential both in vitro and in vivo when compared with BM-MSCs. These results demonstrate the successful isolation of MSCs from human rotator cuffs and encourage the application of RC-MSCs in myogenic regeneration.

Isolation and characterization of mesenchymal stromal cells from human anterior cruciate ligament

BACKGROUND The anterior cruciate ligament (ACL) is one of the most commonly injured ligaments of the knee. Because the torn ACL is always discarded during ACL reconstruction, it may be a potential source for isolating mesenchymal stromal cells (MSC). METHODS To characterize MSC from human ACL, cells were enzymatically released from the ACL of adult human donors and seeded in plastic dishes with serial passages at confluence. At different passages, ACL-derived cells were subjected to in vitro assays to investigate their multilineage potential. Upon treatment, the phenotypes of the cell cultures were analyzed by histo- and immunohistochemistry and semi-quantitative reverse transcription-polymerase chain reaction for the expression of lineage-specific genes. RESULTS Six independent cell lines from individual donors showed diversity in multilineage potential. Interestingly, five of the six lines displayed adipogenic potential, four had osteogenic and adipogenic potential, and only one cell line was tripotent. Both bone marrow (BM)- and ACL-derived MSC expressed marker genes for ligament fibroblasts, whereas the mRNA levels of collagen I and III were more abundant in ACL-derived MSC. DISCUSSION Our study demonstrates that human MSC can be isolated from ACL with diversity in the potential to form bone, fat and cartilage and an increase as compared to BM MSC, in the potential to form ligament fibroblasts.

Imaging evaluation of meniscal injury of the knee joint: A comparative MR imaging and arthroscopic study

To evaluate the efficacy of MR imaging in the diagnosis and classification of meniscal tear of the knee joint, we retrospectively characterized the MR features of 78 meniscal tears in 148 patients according to the Mesgarzadehs criteria. The results showed that the sensitivity and specificity for meniscal tears were 92% and 87%, respectively. Type VI meniscal tear was the most common type, especially in displaced meniscal tear. MR is a reliable diagnostic tool for meniscal tears and associated cruciate ligament injury.

Manipulation under anaesthesia for frozen shoulder in patients with and without non-insulin dependent diabetes mellitus

Manipulation under anaesthesia (MUA) has been used to speed up recovery. However, the outcome of frozen shoulder after MUA in patients with diabetes has not been well documented in the past. A higher prevalence of frozen shoulder has been reported in diabetes mellitus (DM) patients. In this study, we revealed the short- and long-term outcomes for treatment of frozen shoulders by MUA and compared these results in patients with and without non-insulin dependent DM by adjusted Constant score. The scores showed no significant differences between the two groups at both early and late follow-ups. Our results revealed that MUA for frozen shoulders is a simple and noninvasive procedure to improve symptoms and shoulder function within a short period of time. Even though DM is a predisposing factor to frozen shoulder, non-insulin dependent DM alone does not influence both the short- and long-term outcomes of frozen shoulder.

Scale-up of MSC under hypoxic conditions for allogeneic transplantation and enhancing bony regeneration in a rabbit calvarial defect model†

To realize the therapeutic potential of mesenchymal stem cells (MSCs), we aimed to develop a method for isolating and expanding New Zealand rabbit MSCs in a great scale. Rabbit MSCs expanded under hypoxic and normoxic conditions were compared in terms of replication capacity, differentiation potential, and the capacity for allogeneic transplantation in a calvarial defect model. The cells from all tested rabbits were expanded more rapidly when plated at low‐density under hypoxic conditions compared to under normoxic conditions. Moreover, cells expanded under hypoxic conditions increased in the potential of osteoblastic, adipocytic, and chondrocytic differentiation. More importantly, radiographic analysis and micro‐CT measurement of bone volume revealed the hypoxic cells when transplanted in the calvarial defects of another rabbit increased in the ability to repair bone defect compared to the normoxic cells. Six weeks after allogeneic transplantation of hypoxic MSCs, histological analysis revealed a callus spanned the length of the defect, and several bone tissues spotted in the implant. At 12 weeks, new bone had formed throughout the implant. Using BrdU labeling to track the transplanted cells, the hypoxic cells were more detected in the newly formed bone compared to the normoxic cells. For defects treated with allogeneic MSCs, no adverse host response could be detected at any time‐point. In conclusion, we have developed a robust method for isolation and expansion of rabbit MSCs by combining low‐density with hypoxic culture, which can be applied for the design of clinical trials in allogeneic transplantation of MSCs for bone healing.

Comparison of idiopathic, post-trauma and post-surgery frozen shoulder after manipulation under anesthesia.

Manipulation under anesthesia (MUA) has been used to speed up the recovery of frozen shoulder, which is said to be a self-limiting process. We would like to elucidate the short- and long-term results of the treatment of frozen shoulders by manipulation under anesthesia and compare the results of idiopathic, post-trauma and post-surgery frozen shoulders. We applied an adjusted Constant score (Constant score after excluding the 25 points allocated for the assessment of muscle strength) to assess all patients. In our series, 47 cases with 51 frozen shoulders were collected and evaluated retrospectively. The adjusted Constant score at pre-manipulation was on average 22.8±4.9 (10–31) points. The score from the 3-week follow-up was 52.6±9.2 (31–67) points on average. The score from the averaged 82-month follow-up was on average 70.1±6.2 (54–75) points, with 23 shoulders scored for a maximum point number of 75. The score at the early and late follow-ups was significantly lower for the post-surgery group (63.2±6.7) when compared to the other two groups (P<0.001). Our results revealed that manipulation under anestheia is a very simple and noninvasive procedure for shortening the course of an apparently self-limiting disease and can improve shoulder function and symptoms within a short period of time. However, we found less improvement in post-surgery frozen shoulders, especially in residual pain and limited range of motion (ROM), which may be influenced by the initial injury or initial surgery. Although less improvement in pain and ROM was noted, manipulation is still a good and simple way to treat post-surgery frozen shoulders.RésuméLa manipulation sous anesthésie est utilisée pour hâter la récupération des épaules gelées qui sont considérées comme un processus d’auto-limitation. Nous avons étudié les résultats à court et long terme de ces manipulations en comparant les résultats des épaules gelées idiopathiques, post-traumatiques et post-chirugicales avec un score de Constant modifié (en excluant les 25 points alloués à la force musculaire). Nous avons évalué rétrospectivement 51 épaules gelées chez 47 patients. Le score de Constant modifié était avant la manipulation de 22,8±4,9 (10–31). Le score à 3 semaines était de 52,6±9,2 (31–67). Le score à 82 mois de recul moyen était de 70,1±6,2 (54–75), avec 23 épaules au score maximum de 75 points. Le score au recul précoce et tardif était significativement plus faible dans le proupe post-chirurgie (63,2±6,7) que dans les autres groupes (P<0,001). Nos résultats montrent que la manipulation sous anesthésie est une méthode non invasive éfficace pour améliorer rapidement la fonction de l’épaule. Il y a moins d’amélioration dans les cas post-chirurgicaux notamment au niveau des douleurs résiduelles et de l’amplitude de mobilité mais la manipulation reste quand même un bon traitement dans ces cas.

Body mass index and active range of motion exercise treatment after intra-articular injection in adhesive capsulitis.

Background: Adhesive capsulitis is commonly associated with medical diseases such as diabetes mellitus, hyperthyroidism, and obesity. Intra‐articular injection has been used to speed recovery and relieve pain associated with frozen shoulder. In this study, we evaluated and compared the effects of an intra‐articular injection of corticosteroid and lidocaine in the treatment of primary adhesive capsulitis in overweight and normal‐weight patients. Methods: This is a prospective clinical study of patients with adhesive capsulitis, in which the main treatment strategy was an intra‐articular injection of corticosteroid (3 mL) and lidocaine (3 mL). Active range of motion exercise was initiated immediately after the injection and performed four times daily. The evaluation included the recording of a detailed medical and orthopedic history, and the assessment of pain and function by determining the Constant score at baseline (before injection) and every 2 weeks thereafter. Patients were classified as normal weight (body mass index [BMI] < 25 kg/m2) or overweight (BMI ≥ 25 kg/m2). The Constant scores of all patients were compared at 8 weeks after injection. Results: After clinical examinations and radiographic and ultrasonographic studies, 79 patients were treated for adhesive capsulitis between 2010 and 2012. In the normal‐weight group, the mean Constant score increased from 35.4 to 74.6 after 8 weeks, whereas in the overweight group, the mean Constant score increased from 32.0 to 47.2. There was a significant difference in the mean Constant score between the normal‐weight and overweight groups at 8 weeks. Conclusion: Active range of motion exercise after an intra‐articular injection of corticosteroid and lidocaine improved pain and functional outcome at 8 weeks in normal‐weight (BMI < 25 kg/m2) patients with primary adhesive capsulitis. Manipulation under anesthesia may be considered a priority in overweight patients.

Lunocapitate fusion with scaphoid excision for the treatment of scaphoid nonunion advanced collapse or scapho-lunate advanced collapse wrist

Background Four‐corner fusion is an effective procedure to treat advanced degenerative osteoarthritis of scaphoid nonunion advanced collapse or scapho‐lunate advanced collapse wrists. However, lunocapitate fusion, an alternative procedure, shows benefits including less dissection of the soft tissue and also a shorter operation time. We reviewed 10 cases to reveal the complication rates and clinical outcomes of this procedure. Methods We retrospectively reviewed 10 patients with symptomatic scaphoid nonunion advanced collapse or scapho‐lunate advanced collapse wrists who had received lunocapitate fusion with scaphoid excision. The average follow‐up period was 44.5 months (range, 22–68 months). Clinical evaluations were conducted and determined by radiographs, range of motion (flexion–extension), visual analog scale, and Mayo wrist scores. Complications including nonunion and implant migration were recorded. Results Among these patients, eight developed solid radiographic union while the remaining two patients showed bone resorption and implant migration and needed revision surgeries. The visual analog scale was decreased from 5.0 to 1.1, and the flexion–extension arc was increased from 61° to 72.5°. The average Mayo Wrist Score was 70 points. The results showed outcomes similar to those of previous studies. Conclusion Through our investigation and findings, we conclude that lunocapitate fusion can restore a functional and almost pain‐free wrist. Moreover, these results were maintained at follow‐up sessions, with complication rates being similar to those of previous studies. These results conclude a satisfactory therapeutic alternative to four‐corner fusion for advanced degenerative osteoarthritis of wrists.