Li Hsuan Chiu

The effect of type II collagen on MSC osteogenic differentiation and bone defect repair

The function of type II collagen in cartilage is well documented and its importance for long bone development has been implicated. However, the involvement of type II collagen in bone marrow derived mesenchymal stem cell (BMSC) osteogenesis has not been well investigated. This study elucidated the pivotal role of type II collagen in BMSC osteogenesis and its potential application to bone healing. Type II collagen-coated surface was found to accelerate calcium deposition, and the interaction of osteogenic medium-induced BMSCs with type II collagen-coated surface was mainly mediated through integrin α2β1. Exogenous type II collagen directly activated FAK-JNK signaling and resulted in the phosphorylation of RUNX2. In a segmental defect model in rats, type II collagen-HA/TCP-implanted rats showed significant callus formation at the reunion site, and a higher SFI (sciatic function index) scoring as comparing to other groups were also observed at 7, 14, and 21 day post-surgery. Collectively, type II collagen serves as a better modulator during early osteogenic differentiation of BMSCs by facilitating RUNX2 activation through integrin α2β1-FAK-JNK signaling axis, and enhance bone defect repair through an endochondral ossification-like process. These results advance our understanding about the cartilaginous ECM-BMSC interaction, and provide perspective for bone defect repair strategies.

Effects of Low-Intensity Pulsed Ultrasound, Dexamethasone/TGF-β1 and/or BMP-2 on the Transcriptional Expression of Genes in Human Mesenchymal Stem Cells: Chondrogenic vs. Osteogenic Differentiation

The effects of low-intensity pulsed ultrasound (LIPUS) on the differentiation of human mesenchymal stem cells (hMSCs) were investigated in this study. hMSCs were subjected to LIPUS with or without dexamethasone/transforming growth factor-beta1 (TD) or bone morphogenetic protein-2 (BMP-2) and the effects of this treatment were assessed. TD-treated hMSCs exhibited characteristic chondrogenic morphology and increased messenger RNA (mRNA) expression of chondrogenic markers and LIPUS enhanced the chondrogenic differentiation of hMSCs treated with TD. The expression of Runx2, an osteogenic transcription factor was not altered in either TD treatment group; however, a significant increase was detected in the LIPUS only group. The osteogenic appearance exhibited 3 days after LIPUS and/or BMP-2 treatment. Increases in the mRNA expression levels of osteogenic markers, Runx2 and ALP were also detected. There was no additive or altered effect with combined LIPUS and BMP-2 treatment. LIPUS alone can increase osteogenic differentiation of hMSCs and LIPUS enhances TD-mediated chondrogenic differentiation of hMSCs. Clinically, LIPUS may differentially influence bone vs. cartilage repair.

Differential effect of ECM molecules on re-expression of cartilaginous markers in near quiescent human chondrocytes†‡

The limited source of healthy primary chondrocytes restricts the clinical application of tissue engineering for cartilage repair. Therefore, method to maintain or restore the chondrocyte phenotype during in vitro expansion is essential. The objective of this study is to establish the beneficial effect of ECM molecules on restoring the re‐expression of cartilaginous markers in primary human chondrocytes after extensive monolayer expansion. During the course of chondrocyte serial expansion, COL2A1, SOX9, and AGN mRNA expression levels, and GAG accumulation level were reduced significantly in serially passaged cells. Exogenous type II collagen dose‐dependently elevated GAG level and induced the re‐expression of cartilaginous marker mRNAs in P7 chondrocytes. Chondroitin sulfate did not show significant effect on P7 chondrocytes, while hyaluronic acid inhibited the expression of SOX9 and AGN mRNAs. Upon treatment with type II collagen, FAK, ERK1/2, and JNK were activated via phosphorylation in P7 chondrocytes within 15 min. Furthermore, GFOGER integrin blocking peptide, MEK inhibitor and JNK inhibitor, not p38 inhibitor, significantly reduced the type II collagen‐induced GAG deposition level. Finally, in the presence of TGF‐β1 and IGF‐I, P7 chondrocytes cultured in 3D type II collagen matrix exhibited better cartilaginous features than those cells cultured in the type I collagen matrix. In conclusion, type II collagen alone can effectively restore cartilaginous features of expanded P7 human chondrocytes. It is probably mediated via the activation of FAK‐ERK1/2 and FAK‐JNK signaling pathways. The potential application of type II collagen in expanding a scarcity of healthy chondrocytes in vitro for further tissue engineering is implicated. J. Cell. Physiol. 226: 1981–1988, 2011.

Effects of the nanostructure and nanoporosity on bioactive nanohydroxyapatite/reconstituted collagen by electrodeposition

Hydroxyapatite (HA)/collagen composites were reported to induce bony growth. Various methods for preparing HA-based composites have been investigated as potential biomaterials for bone substitutes. However, no method can generate a thick nanoporous HA. A novel bone regenerative nanocomposite consisting of nano-hydroxyapatite (HA), nano-amorphous calcium phosphate (ACP) and reconstituted collagen by electrodeposition was designed in this research. Specimens with and without nanoporosity were evaluated using electrochemical measurements, material analyses, and cell-material interactions. The results showed that reconstituted collagen/nano-(HA and ACP) illustrated a multinanoporous structure and enhanced biocompatibility. Nanocomposite was comprised to nano-(HA and ACP) and reconstituted collagen. The core cell structure was formed during electrodeposition. Nanoporosity and nanostructure were observed as formation of nanocomposite. The nano-(HA and ACP) phases were essentially composed of a nanoporous and nanostructural biocomposite. Reconstituted collagen incorporation with the nanoporous and nanostructural biocomposite significantly facilitated the osteogenic differentiation of mesenchymal stem cells. Reconstituted collagen was covered with nano-(HA and ACP), profoundly impacting the enhancement of biocompatibility on application of implant and tissue engineering. The bioactive nano-HA/reconstituted collagen-induced osteogenic differentiation of mesenchymal stem cells enables to enhance bone growth/repair and osseointegration.

Comparing robotic surgery with conventional laparoscopy and laparotomy for cervical cancer management.

Objective The aim of this study was to compare the outcomes of robotic surgery, laparoscopy, and laparotomy for the surgical treatment of stage IA to IIB cervical cancer. Methods This retrospective study was carried out in a university-affiliated teaching hospital. A total of 100 women with an initial diagnosis of stage IA to IIB cervical cancer, without preoperative brachytherapy or chemotherapy, were included in this study. With selection of the cases, 44 patients received laparotomy surgery, 32 patients received laparoscopic surgery, and 24 patients received robotic surgery. The perioperative parameters measured included operation time, blood loss, transfusion rate, lymph node yield, adhesion score, laparotomy conversion rate, postoperative and 24-hour pain scores, time to full diet resumption, and hospital stay. The perioperative complication and disease-free survival were also evaluated. Results The robotic group showed a shorter operation time, less blood loss, lower transfusion rate, and lower laparotomy conversion rate than the laparoscopic or laparotomy group. As for the postoperative parameters, the robotic group showed reduced postoperative and 24-hour pain scores, shortened length of hospital stay, and decreased time to full diet resumption compared with the other 2 surgical groups. No significant differences were found between the groups in perioperative complication rate or disease-free survival. Conclusions The data suggested that robotic surgery is a feasible and potentially optimal option for the treatment of stage IA to IIB cervical cancer with favorable short-term surgical outcomes.

Comparison of robotic surgery and laparoscopy to perform total hysterectomy with pelvic adhesions or large uterus.

Background: Currently, benefits of robotic surgery in patients with benign gynecological conditions remain unclear. In this study, we compared the surgical outcome of robotic and laparoscopic total hysterectomies and evaluated the feasibility of robotic surgery in cases with pelvic adhesions or large uterus. Materials and Methods: A total of 216 patients receiving total hysterectomy via robotic or laparoscopic approach were included in this study. Of all 216 patients, 88 underwent robotic total hysterectomy and 128 underwent laparoscopic total hysterectomy. All cases were grouped by surgical type, adhesion score, and uterine weight to evaluate the interaction or individual effect to the surgical outcomes. The perioperative parameters, including operation time, blood loss, postoperative pain score, time to full diet resumption, length of hospital stay, conversion rate, and surgery-related complications were compared between the groups. Results: Operation time and blood loss were affected by both surgical type and adhesion score. For cases with severe adhesions (adhesion score greater than 4), robotic surgery was associated with a shortened operation time (113.9 ± 38.4 min versus 164.3 ± 81.4 min, P = 0.007) and reduced blood loss (187.5 ± 148.7 mL versus 385.7 ± 482.6, P=0.044) compared with laparoscopy. Moreover, robotic group showed a lower postoperative pain score than laparoscopic group, as the effect was found to be independent of adhesion score or uterine weight. The grade-II complication rate was also found to be lower in the robotic group. Conclusions: Comparing to laparoscopic approach, robotic surgery is a feasible and potential alternative for performing total hysterectomy with severe adhesions.

Diverse effects of type II collagen on osteogenic and adipogenic differentiation of mesenchymal stem cells

Type II collagen is known to modulate chondrogenesis of mesenchymal stem cells (MSCs). In this study, MSCs from human bone marrow aspirates were used to study the modulating effects of type II collagen on MSC differentiation during the early stages of osteogenesis and adipogenesis. With osteogenic induction, MSCs cultured on the type II collagen‐coated surface showed an enhanced calcium deposition level with increasing mRNA expressions of RUNX2, osteocalcin, and alkaline phosphatase. A synthetic integrin binding peptide, which specifically interacts with the I‐domain of α1β1/α2β1 integrins significantly blocks the mineralization‐enhancing effect of type II collagen. MSCs attached on the type II collagen‐coated plates exhibited expanded cell morphology with increasing spreading area, and the pretreatment of cells with integrin α1β1 or α2β1‐blocking antibody reduced the effect. The phosphorylation levels of FAK, ERK, and JNK significantly increased in the MSCs that attached on the type II collagen‐coated plates. On the contrary, the mineralization‐enhancing effect of type II collagen was diminished by JNK and MEK inhibitors. Furthermore, type II collagen blocked the adipogenic differentiation of MSCs, and this effect is rescued by JNK and MEK inhibitors. In conclusion, type II collagen facilitates osteogenesis and suppresses adipogenesis during early stage MSC differentiation. Such effects are integrin binding‐mediated and conducted through FAK–JNK and/or FAK–ERK signaling cascades. These results inspire a novel strategy encompassing type II collagen in bone tissue engineering. J. Cell. Physiol. 227: 2412–2420, 2012.

A cohort study to evaluate the effectiveness of laparoscopic-guided local injection of etoposide in the management of women with unruptured tubal pregnancy

OBJECTIVE To assess the feasibility of laparoscopic-guided local injection of etoposide or methotrexate (MTX) in the management of unruptured tubal pregnancy and compare the effectiveness of the two regimens. DESIGN Retrospective cohort study. SETTING Medical center. PATIENT(S) Thirty-one women with laparoscopically diagnosed unruptured tubal pregnancy. INTERVENTION(S) A regimen of etoposide 50 mg via laparoscopic-guided local injection (n = 17) compared with a conventional MTX 50 mg regimen (n = 11), after 3 patients were excluded (2 refusals and 1 with salpingostomy). MAIN OUTCOME MEASURE(S) Serial serum β-hCG levels and the success rate in both groups. RESULT(S) General characteristics of the patients were similar in both groups. The overall success rate was 96.4% (27 of 28). The duration between treatment and nadir of serum β-hCG level (<5 mIU/mL) was significantly shorter in the etoposide group than in the MTX group (19.7 ± 13.0 days vs. 33.4 ± 8.1 days). No patient in the etoposide group and only 1 in the MTX group needed reintervention, which led to 100% and 91% success rates for the etoposide and MTX groups, respectively. Three women in the etoposide group had subsequently successful term deliveries. CONCLUSION(S) Both regimens-etoposide 50 mg and MTX 50 mg via laparoscopic-guided local injection-were acceptable in the management of women with unruptured tubal pregnancy because of their similar and high success rates. More studies are needed to confirm this observation.

Comparison of robotic approach, laparoscopic approach and laparotomy in treating epithelial ovarian cancer.

The purpose of this study was to evaluate the feasibility of robotic surgery and compare its surgical outcomes with those of laparoscopic surgery and laparotomy, with regard to performing staging surgery to manage ovarian cancer.

Female sexual dysfunction: definition, classification, and debates.

Sexual dysfunction refers to difficulties that occur during the sexual response cycle that prevent the individual from experiencing satisfaction from sexual activity. It is relatively difficult to estimate the prevalence of female sexual dysfunction (FSD), because the definition and diagnostic criteria are still controversial and under development. These difficulties reveal our insufficient understanding of the basis of FSD. This review was conducted in an effort to deal with this complicated clinical issue, by examining the most updated clinical criteria of FSD under the context of a redefined female sexual response model.