Attachment and Osteogenic Potential of Dental Pulp Stem Cells on Non-Thermal Plasma and UV Light Treated Titanium, Zirconia and Modified PEEK Surfaces.

Ultraviolet (UV) light and non-thermal plasma (NTP) treatment are chairside methods that can efficiently improve the biological aging of implant material surfaces caused by customary storage. However, the behaviors of stem cells on these treated surfaces of the implant are still unclear. This study aimed to investigate the effects of UV light and NTP treated surfaces of titanium, zirconia and modified polyetheretherketone (PEEK, BioHPP) on the attachment and osteogenic potential of human dental pulp stem cells (DPSCs) in vitro. Machined disks were treated using UV light and argon or oxygen NTP for 12 min each. Untreated disks were set as controls. DPSCs were cultured from the wisdom teeth of adults that gave informed consent. After 24 h of incubation, the attachment and viability of cells on surfaces were assessed. Cells were further osteogenically induced, alkaline phosphatase (ALP) activity was detected via a p-Nitrophenyl phosphate assay (day 14 and 21) and mineralization degree was measured using a Calcium Assay kit (day 21). UV light and NTP treated titanium, zirconia and BioHPP surfaces improved the early attachment and viability of DPSCs. ALP activity and mineralization degree of osteoinductive DPSCs were significantly increased on UV light and NTP treated surfaces of titanium, zirconia and also oxygen plasma treated Bio-HPP (p < 0.05). In conclusion, UV light and NTP treatments may improve the attachment of DPSCs on titanium, zirconia and BioHPP surfaces. Osteogenic differentiation of DPSCs can be enhanced on UV light and NTP treated surfaces of titanium and zirconia, as well as on oxygen plasma treated Bio-HPP.

Two simple and inexpensive methods for preparing DNA suitable for digital PCR from a small number of cells in 96-well plates.

BACKGROUND: Although DNA of high quality can be easily prepared from cultured cells with commercially available kits, many studies involve a large number of samples which increases the cost drastically. We optimized two simple and inexpensive methods for preparing DNA suitable for digital PCR from a small number of cells directly from wells of 96-well plates. METHODS: Cells (number: 103 -104 ) were lysed with a Direct PCR® lysis buffer or a 10% Chelex100® solution. The lysates were further purified and concentrated by means of DNA precipitation with a blue-colored glycogen as a carrier. PCR and digital PCR were used to evaluate the efficiency of the two methods. RESULTS: For 1000 cells from one primary culture and two tumor cell lines, DNA was reproducible and obtained with recovery rate (obtained/expected amount of DNA) in the range of 50%-90% as measured by the fluorometer dyes instrument Qubit. Using 8 out of a total of 10 µL DNA solution for 1000 cells, both conventional PCR and digital PCR were successful. For digital PCR, more than 1600 positive droplets were obtained for DNA from 1000 cells using the Direct PCR® method, corresponding to a yield efficiency of approximately 80%. Further reducing the number of cells down to 100 would be possible with 160 positive droplets expected. Both reagents are inexpensive (0.08€/sample). CONCLUSIONS: Two methods are efficient, especially the Direct PCR® reagent-based method provides a simple and inexpensive method for preparing DNA suitable for digital PCR from small number of cells.

Time Dependency of Non-Thermal Oxygen Plasma and Ultraviolet Irradiation on Cellular Attachment and mRNA Expression of Growth Factors in Osteoblasts on Titanium and Zirconia Surfaces.

Ultraviolet (UV) light and non-thermal plasma (NTP) are promising chair-side surface treatment methods to overcome the time-dependent aging of dental implant surfaces. After showing the efficiency of UV light and NTP treatment in restoring the biological activity of titanium and zirconia surfaces in vitro, the objective of this study was to define appropriate processing times for clinical use. Titanium and zirconia disks were treated by UV light and non-thermal oxygen plasma with increasing duration. Non-treated disks were set as controls. Murine osteoblast-like cells (MC3T3-E1) were seeded onto the treated or non-treated disks. After 2 and 24 h of incubation, the viability of cells on surfaces was assessed using an MTS assay. mRNA expression of vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) were assessed using real-time reverse transcription polymerase chain reaction analysis. Cellular morphology and attachment were observed using confocal microscopy. The viability of MC3T3-E1 was significantly increased in 12 min UV-light treated and 1 min oxygen NTP treated groups. VEGF relative expression reached the highest levels on 12 min UV-light and 1 min NTP treated surfaces of both disks. The highest levels of HGF relative expression were reached on 12 min UV light treated zirconia surfaces. However, cells on 12 and 16 min UV-light and NTP treated surfaces of both materials had a more widely spread cytoskeleton compared to control groups. Twelve min UV-light and one min non-thermal oxygen plasma treatment on titanium and zirconia may be the favored times in terms of increasing the viability, mRNA expression of growth factors and cellular attachment in MC3T3-E1 cells.

Clinical analysis of the preoperative condition and operative prognosis of post-cesarean section scar diverticulum: a case series.

Objectives Post-cesarean section scar diverticulum (PCSD) is a long-term sequela of cesarean section (CS). The aim of this study was to evaluate the clinical utility of PCSD scoring criteria, and also retrospectively investigate the efficacy and fertility of two different surgical methods in 304 patients with PCSD. Methods A total of 304 PCSD patients who underwent hysteroscopy or combined hysteroscopy and laparoscopy (referred to as laparoscopy) in our hospital from 2016 to 2018 were retrospectively analyzed. Preoperative condition was analyzed by the PCSD scoring criteria and its influencing factors were explored. The efficacy, its influencing factors and pregnancy success rate of the two different surgical methods on PCSD was also analyzed after 6- and 12-months follow-up. Results PCSD was more severe (high score) in patients who experienced caesarean section with one of the following conditions: age >30 years old, without medical indications or retroflexed uterus. The postoperative efficacy of patients subjected to hysteroscopy or laparoscopy was 81.25 and 89.47% (after 6 months), and 79.53 and 87.50% (after 12 months), respectively. Hysteroscopic surgery was better for PCSD patients who had fewer CS and thicker residual muscle layer and worse for PCSD patients with a longer distance of incision defect to the end of the cervix. Postoperative fertilization showed that pregnancy success rate of patients subjected to hysteroscopy or laparoscopy was 56.2 and 50%, respectively. Conclusions The PCSD scoring is an effective method for assessing the severity of PCSD, and hysteroscopy and laparoscopy are effective modalities for PCSD. Hysteroscopy is also an option for patients with fertility needs.

Hysteroscopic treatment of cesarean scar defect.

OBJECTIVE: To investigate the effect of hysteroscopic surgery on the outcomes of obstetrics and gynecology among patients with cesarean section diverticulum. METHODS: Ninety-nine infertile patients with cesarean section diverticulum received hysteroscopic treatment and were retrospectively analyzed. Patients were followed for 1 year. RESULTS: The study included ninety-nine symptomatic patients with cesarean section diverticulum. After surgery, the menstrual periods of patients were improved from 11.15 ± 4.44 to 7.69 ± 2.85 days. Forty-seven (47/99) women became pregnant after surgery. The number of patients who became pregnant with an anteflexion uterus after hysteroscopic surgery is 32 (32/57), and the number of women who became pregnant with a retroflexion uterus is 15 (15/42). CONCLUSION: Hysteroscopic surgery could improve the PCSD-associated prolonged menstrual bleeding, and satisfactory obstetrical outcomes could be achieved by the surgery treatment in women with cesarean defect.

Propranolol Specifically Suppresses the Viability of Tumorous Schwann Cells Derived from Plexiform Neurofibromas In Vitro.

BACKGROUND/AIM: Plexiform neurofibromas (PNFs) are benign tumors of the periph eral nerves sheath, which can damage neighboring organs, impair functions, cause pain and serious maxillofacial disfigurement, and have a high risk of malignant transformation. Complete resection is usually not possible since PNFs often extend through multiple layers of tissue. Therefore, it is necessary and beneficial to find a reasonable drug treatment for PNFs. Propranolol-treatment is the first-line therapy for infantile hemangiomas and the side effects are reversible and mostly benign. The present study aimed to examine the possible effect of propranolol for suppressing PNFs in vitro. MATERIALS AND METHODS: Paired primary Schwann-cell-rich cultures and fibroblast-rich cultures were obtained from 4 PNFs of unrelated patients. Human Schwann cells (HSCs) were used as the control. These cultures were treated with propranolol for 7 days at concentrations up to 150 µM. Cells were then measured for their viability and immune-stained with S100 to label the tumorous Schwann cells. RESULTS: Propranolol inhibited the viability of the tumorous Schwann cells in a dose-dependent manner, while did not substantially suppress viability of the non-tumorous fibroblasts derived from the same PNFs. CONCLUSION: Propranolol may provide a treatment option for suppressing the growth of PNFs.

[Giant solitary fibrous tumor of vagina: A case report and literature review].

We reported a case of giant solitary fibrous tumor of vagina and reviewed literature. The clinical features, diagnosis, and treatment schemes for the disease were summarized to improve the understanding of the disease. An elder female patient came to the Third Xiangya Hospital, Central South University, because of abdominal distention and pain for 5 days after menopause for 9 years. The patient was diagnosed as a solitary fibrous tumor of vagina by pathology and immunohistochemistry after complete resection. The tumor size of the patient was the largest according to reported literature, and the tumor recurred 10 months after surgery. The strong positive expression of CD34 and high Ki-67 proliferation index in tumor immunohistochemistry indicate that the prognosis of patients will be poor.