[ laboratory study on human jaw osteoblastic stem cells culturing]

The goal of this study was to evaluate different methods of cultivating human bone cells. Five periosteal and bone specimens obtained from the human jaw were divided into small pieces in the laboratory. After the addition of trypsin and collagenase enzymes and releasing of cells, three primary periostal, endosteal, and bony chips cells were prepared. After passing the required time for the growth of specimens, lamella were prepared and stained with alkaline phosphatase [ALP] in order to determine ALP positive cells. Mean time of cellular growth was 23 days. Human bone cells have the capability of being cultured under special sterile laboratory conditions and three dimensional culturing of them can be used for reconstruction of maxillofacial region defects

In vitro survival rate of bovine oocytes following vitrification in glass capillary micropipette [GCM]

The purpose of this study was to evaluate the use of glass capillary micropipette [GCM] as a vessel for vitrification of bovine oocytes. Cumulus-oocyte complexes [COCs] were obtained from slaughter-house and washed 5 to 6 times in the washing medium [TCM-199 + 20% FBS] and randomly assigned to treatment and control group. In the first step of vitrification, COCs were exposed to first vitrification solution [VS1] [10% ethylene glycol [EG], 10% DMSO in holding medium [TCM-l99 + 10% FBS: HM]] for I min at room temperature and then placed in VS2 solution [20% EG, 20% DMSO in HM] for 25 sec and immediately were loaded into the GCM vessel. The filled portion of GCM vessels were placed in liquid nitrogen [LN [2]] for 3 to 5 sec and then completely immersed and stored there. The oocytes were thawed by immersing the capillary end of the straw in 1 ml of 0.25 M sucrose in HM and gently expelling the contents. After 1 min the oocytes were transferred into 100 micro l of 0.15 M sucrose in HM for another 5 mm and then washed with HM twice. For examining the in vitro developmental potential of vitrified-warmed oocytes, the oocytes were placed in 50 micro 1 droplet of maturation medium [TCM-l99 + 10% FBS - 10 IU/ml PMSG + 15 IU/ml HCG] covered with paraffin oil in a CO [2] incubator at 38.5°C for 24 hrs. A high proportion of morphologically normal oocytes [90%] was recovered after vitrification-warming. The percentage of live oocytes after 24 hrs when tested with trypan blue in GCM group was 85.18%, significantly did not differ from control group [90%]. The proportion of oocytes which were found to have undergone nuclear maturation did not show statistical difference between the control and GCM group [61.29% vs 40%, respectively]. The results of present study demonstrated that vitrification of immature bovine oocytes in the GCM vessels and EG + DMSO solution have high survival rate

[Effects of Morphine dependency on the ovarian folliculogenesis following superovulation in mice]

Opioids may affect hypothalamic GnRH secretion and Hypothalamic-Pituitary- Gonad axis, resulting in reproductive disturbances. Current study investigates the effects of morphine on structure of ovary following superovulalion through morphologic/morphometric studies. Twelve young female NMRI mice were allocated into treatment and control groups. Treatment group received oral morphine at final dose of 0.4mg/ml for 21 days. Physical dependency was proved by injection of naloxone [2mg/kg ip]. The mice were superovulated by 10 iu PMSG [ip] and 48 hours later were sacrificed by cervical dislocation. Ovaries were removed and H and E staining was done. Every 10[th] serial section, which represents nonrandom 10 percent sample was counted. Follicles were classified into small, growing, antral and atretic according to the diameter and number of follicular cell layers surrounding oocytes. The volume and the weight of ovaries were recorded. In addition, the diameter of the antral follicles and oocytes was carefully measured by a calibrated oculometer. The volume and the weight of ovaries showed no significant alterations in the two groups. The proportion of small and atretic follicles was statistically different in treatment and control groups [P<0.001]. According to our data, oral morphine did not alter the volume and the weight of the ovary. However, folliculogenesis was moderately affected by morphine and following superovulation the behavior of ovaries in the treatment group is comparable to the control group