Dah-Ching Ding

Human Infrapatellar Fat Pad-Derived Stromal Cells Have More Potent Differentiation Capacity Than Other Mesenchymal Cells and Can Be Enhanced by Hyaluronan.

The microenvironment plays an important role in the homing in and differentiation of stem cells to repair injured tissue. Infrapatellar fat pad stromal cells (IFPSCs) are a promising source of such cells for the repair of articular injury-induced degeneration. This study investigated the chemotaxis of IFPSCs to chondrocytes and the effect of hyaluronan (HA) on the biological and regenerative properties of IFPSCs. The IFPSCs were obtained from patients undergoing arthroscopy and cultured via a standard 2-week culture protocol that yielded more than 10 million cells on passage 3. The results showed that the IFPSCs had a higher capacity for chondrogenic differentiation than mesenchymal cells from body fat, bone marrow, and Whartons jelly of the umbilical cord. The IFPSCs cultured on 25% or 50% HA showed better osteogenic and adipogenic capabilities than those without HA or with 75% HA (p < 0.001). Cultures of the IFPSCs on 25% HA had a fourfold increase in chondrogenic differentiation compared to cultures without HA, which was better than with 50% and 75% HA (p < 0.05). Cell proliferation was not affected by the presence of HA. In conclusion, IFPSCs have a strong potential for chondrogenic regeneration, which can even be augmented in a 25% HA microenvironment.

Scar Endometriosis at the Site of Cesarean Section

OBJECTIVE Scar endometriosis is a rare condition. We report a case of scar endometriosis occurring at the site of an old cesarean section scar. CASE REPORT A 29-year-old multiparous woman complained of painful sensation during menstruation for 2 years that occurred at the site of her cesarean section surgical scar. On examination, there was a firm nodule measuring 2 x 1.5 cm in size at the mid-point of the scar. In view of the possibility of scar endometriosis, the mass was completely excised. Pathologic findings were compatible with scar endometriosis. Postoperatively, danazol was prescribed to prevent recurrence. CONCLUSION A surgical scar becoming painful and swollen during menstruation is the classic symptom of scar endometriosis. Causes include iatrogenic transplantation of endometrium to the surgical wound. Surgical excision is the main treatment. Postoperative GnRH-agonist or danazol may be prescribed to patients with scar endometriosis.

Characterization of HLA-G and Related Immunosuppressive Effects in Human Umbilical Cord Stroma-Derived Stem Cells:

Mesenchymal stem cells (MSCs) and especially those derived from fetal tissues exert a potent immunosuppressive effect that can be enhanced under inflammatory conditions. This study aimed to explore the immunosuppressive properties of human umbilical cord mesenchymal stem cells (HUCMSCs). We found that HLA-G, the nonclassical HLA allele with strong immune-inhibitory properties, was much more expressed on the HUCMSCs than on MSCs of other origins. Flow cytometry revealed that 90.8% of the HUCMSCs expressed HLA-G. RT-PCR revealed expression of HLA-G1, HLA-G5, and HLA-G7 in all of four HUCMSC lines. In a mixed lymphocyte reaction assay, the HUCMSCs inhibited the proliferation of lymphocytes by 35 ± 3% and could be reversed by treatment with an HLA-G blocking antibody. Upon coculture with the HUCMSCs, peripheral blood mononuclear cells expressed lower levels of proinflammatory mediators such as IL-6, TNF-α, and VEGF-α. This immunosuppressive effect was enhanced when the HUCMSCs were pretreated with IFN-γ, such that the expression of HLA-G was highly activated and HLA-DR diminished. The same phenomenon was not observed in MSCs derived from bone marrow or the placenta. In a xenograft rejection assay, the HUCMSCs survived in immunocompetent mice, whereas primary fibroblasts did not survive. This study confirms the HLA-G-related immunosuppressive property of HUCMSCs, which is more potent than MSCs of other origin. A good tolerance of this mesenchymal stem cell in allogeneic transplantation can thus be anticipated.

Long-term persistence of human papillomavirus in environments.

OBJECTIVE The possibility of its indirect transmission of human papillomavirus (HPV) via formites has been widely raised but with no biological proof. This study explored the durability of HPV16 pseudoviruses and native viruses in different environmental contamination scenarios. METHODS Pseudoviruses were mixed with PBS, cervico-vaginal secretion (CVS), or serum to simulate contamination by genital warts, vaginal discharge or menstruation, respectively, and subjected to in-vitro cell infection assay. The integrity of native HPV16 from CVS of infected women was detected by conformation-specific antibody. RESULTS In viruses exposed to PBS, a persistent infectivity of 30% was noted for at least 7 days. A similar persistence but lower (18%) infectivity was noted in those exposed to CVS. In serum-containing medium, the infection ratio rose initially, remained stable for three more days then rapidly decreased thereafter. Upon desiccation, infectivity was persistently low (10%). Finally, intact native HPV was detectable after 5 days of environmental exposure. CONCLUSION This study demonstrated the high environmental survivability of HPV. However, survivability was lower in viruses exposed to CVS or desiccation.

Mesenchymal Stem Cells and Their Clinical Applications in Osteoarthritis.

Osteoarthritis is a chronic degenerative joint disorder characterized by articular cartilage destruction and osteophyte formation. Chondrocytes in the matrix have a relatively slow turnover rate, and the tissue itself lacks a blood supply to support repair and remodeling. Researchers have evaluated the effectiveness of stem cell therapy and tissue engineering for treating osteoarthritis. All sources of stem cells, including embryonic, induced pluripotent, fetal, and adult stem cells, have potential use in stem cell therapy, which provides a permanent biological solution. Mesenchymal stem cells (MSCs) isolated from bone marrow, adipose tissue, and umbilical cord show considerable promise for use in cartilage repair. MSCs can be sourced from any or all joint tissues and can modulate the immune response. Additionally, MSCs can directly differentiate into chondrocytes under appropriate signal transduction. They also have immunosuppressive and anti-inflammatory paracrine effects. This article reviews the current clinical applications of MSCs and future directions of research in osteoarthritis.

Mutagenic, surviving and tumorigenic effects of follicular fluid in the context of p53 loss: initiation of fimbria carcinogenesis.

Ovulation is the strongest risk factor for ovarian high-grade serous carcinoma (HGSC) that largely originates from the fallopian tube fimbriae and always carries loss-of-function mutations of TP53 in both early and late lesions. Mature ovarian follicle contains high level of reactive oxygen species (ROS). When released from ovulation, follicular fluid (FF) bathes the fimbriae and may lead to DNA double-strand break (DSB) and neoplastic transformation. In this study, we examined the mutagenic and tumorigenic activities of human pre-ovulatory FFs. A subset (6/11) of FFs was found with high levels of ROS whereas the antioxidant capacities were indifferent. These ROS(high) FFs induced intracellular ROS and DSBs in the secretory cell population of fimbriae epithelium. When p53 and Rb were turned down, the FF-exposed secretory cells overcame apoptosis and expanded the population carrying ROS and DSB. The cancer initiation and promotion effects of FF were further recapitulated in Trp53 (-/-) mice. When introduced into the mammary fat pad, ROS(high) but not ROS(low) FFs induced early-onset B-cell lymphoma. Cotreatment with physiological concentration of melatonin, a potent antioxidant, ameliorated the mutagenic and tumorigenic effect of ROS(high) FF in vitro and in vivo. The study revealed ROS and mitogens in mature ovarian follicles could initiate the transformation of fimbria epithelium in the context of p53 loss and melatonin is a potent preventive agent.

Repair of Uterine Rupture in Twin Gestation after Laparoscopic Cornual Resection

Spontaneous uterine rupture in the course of pregnancy is a rare event that usually occurs in a scarred uterus. The event occurs mostly during the intrapartum period and is potentially catastrophic for both mother and fetus. We report a case of 2-cm cornual rupture in a pregnant woman at 13 weeks twin gestation with previous history of cornual pregnancy successfully managed via laparoscopy. Sudden onset of abdominal pain and vaginal bleeding was noted first. Physical examination revealed stable vital signs, lower abdominal tenderness, and mild rebounding pain. Pelvic ultrasonography revealed twin pregnancy at 13 weeks with extrauterine saccular structure 6 cm in diameter located on the left fundus and contiguous with an intrauterine oligohydramnics twin. Exploratory laparotomy was promptly performed, and a small rupture about 2 cm in diameter was observed on the upper portion of the left fundus, the site of a previous laparoscopic cornual resection scar. A protruding amniotic sac of about 6 cm diameter and containing some part of the umbilical cord was seen. The uterine rupture site was repaired directly after aspiration of amniotic fluid from the protruding sac. After surgery, the patient received antibiotics, 17-OH-progesterone for potential rupture of membranes and prematurity. Tocolysis with Ritodrine for irregular uterine contractions was given at 22 weeks gestation. Steroids were given at 24 weeks gestation. The pregnancy ended with a successful delivery by cesarean section because of uncontrollable uterine contractions at 30 5/7 weeks gestation. In conclusion, although termination of pregnancy would normally be recommended when uterine rupture occurs, a different approach to management may now be accepted.

Enhanced differentiation and clonogenicity of human endometrial polyp stem cells

Endometrial polyps arise from endometrial overgrowth and may cause intermenstrual bleeding, irregular bleeding, and menorrhagia. In this study, endometrial polyps were harvested from hysterectomized specimens from 6 female patients not on hormone therapy. Endometrial polyp mesenchymal stem cells (EPMSCs) were isolated and characterized. Selected cells were spindle-shaped, and expressed surface markers CD90 and CD146. The EPMSCs proliferated actively in vitro. A colony-forming study demonstrates that EPMSCs had a colony-generating capacity. When cultured in a defined medium, EPMSCs can differentiate to osteoblast-, adipocyte-, and neuron-like cells. No telomerase reverse transcriptase (TERT) expression was noted. Experimental results demonstrate that EPMSCs are a population of mesenchymal progenitor cells existing in human endometrial polyps that are capable of proliferation, differentiation, and colonogenicity exceeding that of bone marrow stem cells and endometrial stromal cells. These EPMSCs may be an alternative resource of adult stem cells for future regenerative therapy.

Haemoglobin in pelvic fluid rescues fallopian tube epithelial cells from ROS stress and apoptosis

Fallopian tube fimbrial epithelium is considered to be the major site of origin of ovarian high‐grade serous carcinoma, with p53 loss being the earliest and universal change. We previously reported that reactive oxygen species (ROS) in the ovulatory follicular fluids (FFs) are mutagenic and cytotoxic to fimbrial epithelial cells, which are bathed in the peritoneal fluid mixed with FFs. Here, we observed that ferryl haemoglobin (Hb), which was abundantly present in ovulatory FFs and pelvic peritoneal fluids, could rescue p53‐deficient immortalized fimbrial epithelial (FE25) cells and oviduct epithelial cells from Trp53‐null mice from lethal ovulatory ROS stress. Ferryl Hb and FF containing high Hb levels protected FE25 cells from apoptosis, mainly by consuming extracellular ROS and reducing NADPH oxidase‐mediated cell death. The remaining extracellular ROS could still induce DNA double‐strand breaks in the fimbrial epithelial cells. Our study revealed that ferryl Hb in peritoneal fluid rescued ROS‐stressed, DNA‐damaged fimbrial epithelial cells from death, and suggested that peritoneal blood from various sources may contribute to the ovulation‐induced transformation of Fallopian tube epithelium. Copyright

Haemoglobin in pelvic fluid rescues Fallopian tube epithelial cells from reactive oxygen species stress and apoptosis

Fallopian tube fimbrial epithelium is considered to be the major site of origin of ovarian high‐grade serous carcinoma, with p53 loss being the earliest and universal change. We previously reported that reactive oxygen species (ROS) in the ovulatory follicular fluids (FFs) are mutagenic and cytotoxic to fimbrial epithelial cells, which are bathed in the peritoneal fluid mixed with FFs. Here, we observed that ferryl haemoglobin (Hb), which was abundantly present in ovulatory FFs and pelvic peritoneal fluids, could rescue p53‐deficient immortalized fimbrial epithelial (FE25) cells and oviduct epithelial cells from Trp53‐null mice from lethal ovulatory ROS stress. Ferryl Hb and FF containing high Hb levels protected FE25 cells from apoptosis, mainly by consuming extracellular ROS and reducing NADPH oxidase‐mediated cell death. The remaining extracellular ROS could still induce DNA double‐strand breaks in the fimbrial epithelial cells. Our study revealed that ferryl Hb in peritoneal fluid rescued ROS‐stressed, DNA‐damaged fimbrial epithelial cells from death, and suggested that peritoneal blood from various sources may contribute to the ovulation‐induced transformation of Fallopian tube epithelium. Copyright