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Biomedical subjects

D W Han

Publications and source records attributed to D W Han.

17 recordsLinked to original sources

A bone replaceable artificial bone substitute: cytotoxicity, cell adhesion, proliferation, and alkaline phosphatase activity.

Cellular toxicity, cell adhesion and proliferation, and alkaline phosphatase (ALP) activity were investigated for an artificial bone substitute composed of heated carbonate apatite (CAp) and Type I atelocollagen (AtCol) extracted from bovine tail skins (88/12 in %wt/wt). To enhance the intramolecular crosslinking between collagen molecules, the CAp-AtCol substitutes were irradiated by ultraviolet rays (wave length 254 nm) at 4 degrees C for 4 h or vacuum dried at 150 degrees C for 2 h. Cytotoxicity tests by a direct contact method and an extract dilution method revealed that the CAp-AtCol substitutes were cytocompatible for balb 3T3 fibroblasts. Osteoblast adhesion studies demonstrated that the substitute disks composed of 980 degrees C-heated CAp and AtCol were significantly more adhesive for osteoblasts than those of 1,200 degrees C-sintered CAp and AtCol (p < 0.05). Proliferation studies showed that the number of osteoblasts grown in the media containing substitutes of 980 degrees C-heated CAp and AtCol was statistically higher than grown in those of 1,200 degrees C-sintered CAp and AtCol after 5 days (p < 0.05). It was found that osteoblasts grown in the substitutes of 980 degrees C-heated CAp and AtCol only expressed similar ALP activity to the controls. These results suggested that the substitutes consisting of 980 degrees C-heated CAp and AtCol show more favorable interactions with osteoblasts than those of 1,200 degrees C-sintered CAp and AtCol.

3T3 Cells↗

A bone replaceable artificial bone substitute: osteoinduction by combining with bone inducing agent.

Bone inducing agent (BIA) isolated from Saos-2 human osteosarcoma cells was added to an artificial bone substitute composed of 980 degrees C-heated carbonate apatite (CAp) and Type I atelocollagen (AtCol) extracted from bovine tail skins (88/12 in wt/wt %), and a CAp-AtCol-BIA substitute was prepared as an osteoinductive bone substitute. Rat calvaria osteoblasts treated by the isolated BIA demonstrated significantly increased alkaline phosphatase (ALP) activity after 3 days (p < 0.05). In vitro cell attachment and proliferation and ALP activity were investigated for the bone substitute combined with BIA. Osteoblasts cultured onto the surface of the CAp-AtCol-BIA substitute demonstrated remarkable morphological changes such as radial spreading, flattening, and projecting filopodia after 5 days. In comparison with the substitute without BIA, osteoblasts grown in the BIA-combined CAp-AtCol substitute expressed significantly increased proliferation and ALP activity, respectively (p < 0.05). Both the substitutes combined with and without BIA were implanted into artificial defects created in rabbit radii. After 4 weeks, the CAp-AtCol-BIA substitute implanted lesion was completely replaced by regenerated host bone in radiological observation whereas the substitute without BIA was partially resorbed. No histologic abnormalities appeared in the substitute either with or without BIA.

Alkaline Phosphatase↗

A novel in vitro assessment of tissue valve calcification by a continuous flow type method.

A dynamic flow type testing to study calcification was self-designed to investigate calcification in bioprosthetic heart valves. The apparatus consists of a container into which leaflets from a porcine aortic valve are placed, a chamber that contains calcium solution, and a peristaltic pump that provides a continuous supply of the solution toward the container. Efficacy of the apparatus was compared with the conventional batch type calcification testing at 37 degrees C through measuring the amount of calcium and phosphate deposited by inductively coupled plasma (ICP) and scanning electron microscope (SEM). After 14 days, calcium levels detected from the calcified deposit on leaflets were 470.4 +/- 37.0 microg/cm3 in the flow type testing whereas in the batch type testing levels were 81.0 +/- 6.7 microg/cm3. Though the calcium level on the leaflet increased as the exposure time to calcium solution increased in both testings, the rate and the tendency of calcification could be assessed very rapidly by flow type testing in comparison with batch type testing. [Ca]/[P] molar ratio decreased over time, and after 14 days, the ratio was close to 1.83 +/- 0.18 in the flow type testing. The ratio could not be determined in the batch type testing because the deposit was too small to assess. The descending rate of [Ca]/[P] molar ratio demonstrates that deposited calcium-complex at the earliest stage may interact with inorganic phosphate ions to create a calcified deposit mineral precursor. This in vitro dynamic flow type calcification testing was a favorable tool for rapid investigation of calcification.

Aortic Valve↗

A bone replaceable artificial bone substitute: morphological and physiochemical characterizations.

A composite material consisting of carbonate apatite (CAp) and type I atelocollagen (AtCol) (88/12 in wt/wt%) was designed for use as an artificial bone substitute. CAp was synthesized at 58 degrees C by a solution-precipitation method and then heated at either 980 degrees C or 1,200 degrees C. In this study, type I AtCol was purified from bovine tail skins. A CAp-AtCol mixture was prepared by centirfugation and condensed into composite rods or disks. The scanning electron-microscopic (SEM) characterization indicated that the CAp synthesized at 58 degrees C displayed a crystallinity similar to that of natural bone and had a high porosity (mean pore size: about 3-10 microns in diameter). SEM also revealed that the CAp heated at 980 degrees C was more porous than that sintered at 1,200 degrees C, and the 1,200 degrees C-heated particles were more uniformly encapsulated by the AtCol fibers than the 980 degrees C-heated ones. A Fourier transformed-infrared spectroscopic analysis showed that the bands characteristic of carbonate ions were clearly observed in the 58 degrees C-synthesized CAp. To enhance the intramolecular cross-linking between the collagen molecules, CAp-AtCol composites were irradiated by ultraviolet (UV) ray (wave length 254 nm) for 4 hours or vacuum-dried at 150 degrees C for 2 hours. Compared to the non cross-linked composites, the UV-irradiated or dehydrothermally cross-linked composites showed significantly (p < 0.05) low collagen degradation and swelling ratio. Preliminary mechanical data demonstrated that the compressive strengths of the CAp-AtCol composites were higher than the values reported for bone.

Animals↗

Viability and enzymatic activity of cryopreserved porcine heart valve.

Fibroblast viability of a natural tissue valve for replacing a defective heart valve through allograft or xenograft has been suggested to affect its clinical durability. In this study, the cell viability and enzymatic activity of porcine heart valve leaflets were examined in regard to concerning to the preservation process [variable warm ischemic time (WIT), cold ischemic time (CIT), and cryopreservation]. Porcine heart enblocs were obtained and valve dissection was performed after 2, 12, 24, or 36 hours, in respective groups A, B, C, and D, as WIT. Each group was stored for 24 hours as CIT and cryopreserved. Leaflets were dissected from a valved conduit after each process, and cell viability and enzymatic activity in the leaflet were investigated using trypan blue staining and API ZYM kits. WIT extension significantly decreased fibroblast viability (p < 0.05, 92.25 +/- 2.7% at 2 hours, 84.9 +/- 6.7% at 12 hours, 57.0 +/- 10.2% at 24 hours, 55.9 +/- 7.9% at 36 hours), while CIT for 24 hours was also influenced significantly (p < 0.05), whereas cryopreservation demonstrated no effect on cellular viability. In enzyme activity observation, several enzymes related to lipid or nucleotide degradation (esterase, esterase lipase, particularly phosphatase, phosphohydrolase) were remarkably changed following the valve-fabrication process. After 24 hours CIT, these enzymatic activities in groups B, C and D significantly increased, but the activities decreased after cryopreservation. Particularly, both the viability and enzymatic activity showed remarkable changes after CIT in group B (WIT = 12 hours). These results suggest that WIT is more important than CIT in maintaining viability of the valve, and that completing all the cryopreservation process within 12 hours after acquisition is recommended.

Animals↗

A preliminary study of steroid reproductive hormones in human hair.

Nowadays research and clinical studies of human reproductive endocrinology are generally carried out using human blood reproductive hormone assays. However the acquisition of human blood samples has some shortcomings. In search of new approaches, we paid attention to the fact that progesterone can be detected in cow's hair. Consequently we investigated whether or not steroid hormones are measurable in human hair. The results showed that the levels of steroid hormones in hair are not affected by shampoo and do not significantly vary between different segments of hair (i.e. top, middle and basal segments). The menstrual estradiol and progesterone rhythm of female hair is similar to that of female serum. The ratio of hair estradiol to serum estradiol in the female is 41.2% and that of hair progesterone to serum progesterone is 59.0%; the ratio of hair testosterone to serum testosterone in male is 116%. There are significant correlations between hair and serum steroid hormones of healthy human adult: gamma (estradiol) = 0.395 (n = 20), p < 0.05; gamma (progesterone) = 0.440 (n = 22), p < 0.025 and gamma (testosterone) = 0.395 (n = 25), p < 0.05.

Adult↗

Separation and characterization of polyethylene wear debris from synovial fluid and tissue samples of revised knee replacements.

A study was made of in vivo-generated polyethylene wear particles as separated from synovial fluid samples and from tissue samples surrounding total knee arthroplasty. A comparison of particle size and morphology between the two particle groups was made to assess any effects of selective tissue capture, and macrophage encapsulation and digestion. In addition, a Raman spectroscopy technique was evaluated that enables positive identification of individual wear particles. The particles of the same size range found in the synovial fluid and tissue samples exhibited a comparable morphology. Notably, submicron-sized debris was present in both the synovial fluid and tissue samples surrounding knees with osteolysis. The novel micro-Raman analysis of individual particles was successful in the categorizing of wear debris as polyethylene or nonpolyethylene.

Aged↗

[Study on follicle growth, ovulation and morphological changes of endometrium by ultrasonic and biochemical measurements].

Daily growth of the follicles before ovulation and the changes of the endometrium after ovulation were recorded in 16 spontaneous menstrual cycles by the ultrasonic and biochemical measurements. The mean diameter of the dominant follicle was 20 mm before ovulation, the mean volume 3.0 ml, and the growth rate of the follicles 1-3 mm/24 h. Ovulation occurred within 24 h of the luteinizing hormone peak and within 48 hours of the blood estrogen peak. The fact that the blood progesterone levels were higher on the day of the LH peak indicated that luteinization of the dominant follicles had already occurred prior to ovulation. Sonographic criteria of the endometrical tissue were obtained after the serial observation. According to the different sonographic appearances, the secretory phase of the endometrial tissue was divided into the early secretory phase, the middle secretory phase and the late secretory phase. The sonographic characterization of the endometrial tissue in the different phases as well as the thickness of the endometrium during the cycles were described. The clinical usefulness of the criteria of the different phases was to evaluate the subsequent luteal function, and facilitate the clinical management of the infertile women. The study confirms that ultrasound can provide a reliable measure in monitoring the follicular growth and ovulation, and observing the morphological changes of the endometrial tissue during the secretory phase. Thus the in vivo differentiation of the endometrial tissue during the secretory phase could be studied non-invasively by means of the ultrasound tissue characterization.

Adult↗