PURPOSE: In the autologous fat injection, the centrifugation is useful for the refinement of harvested fat. As it can be an injury to the fat cell, we studied the fat cell viability with the change of centrifugation velocity and centrifugation time in order to get the limits of centrifugation velocity and centrifugation time. METHODS: We used the Colman System in 8 patients. We handled the control group with no centrifugation, group I with the centrifugation with 1500rpm for 1 minute, group II with 1500 rpm for 3 minutes, group III with 1500rpm for 5 minutes, group IV with 3000rpm for 1 minute, group V with 3000rpm for 3 minutes, group VI with 3000rpm for 5 minutes, group VII with 5000rpm for 1 minute, group VIII with 5000rpm for 3 minutes, group IX with 5000rpm for 5 minutes. We used the collagenase to separate the fat tissue. We had evaluated the fat cell viability by checking survival cell counts. RESULTS: There was no significance in group I, II, IV, V, but there was significant difference in group III, VI, VII, VIII, IX. CONCLUSION: The centrifugation with 3000rpm for 3 minutes is recommendable.
Adipocytes* ; Cell Count ; Centrifugation* ; Collagenases ; Humans

Keratocyne(R) is the complex of cystine and vitamine B6. Cystine is collagenase inhibitor and vitamine B6 is essential to the metabolism of cystine. We administered Keratocyne(R) orally combined with specific topical therapy to investigate its healing effect on various corneal diseases. The results were as follows: 1. The cases which improved after Keratocyne(R) administration were 15 patients out of 22 patients. 2. No untoward side effects were observed. We concluded that Keratocyne(R) was effective on certain corneal diseases which released collagenase.
Collagenases ; Corneal Diseases* ; Cystine ; Humans ; Metabolism ; Vitamins

The pathogenesis of scleroderma has not been completely delineated, but it is suggested that increased collagen expression in fibroblast from sclerotic skin lesions may be an important factor contributing to collagen accumulation. PUVA therapy has been reported to be effective in the treatment of localized scleroderma. Its mechanism of action appears to be the stimulation of collagenase production by dermal fibroblasts. We report a case of localized scleroderma improved with systemic PUVA therapy.
Collagen ; Collagenases ; Fibroblasts ; PUVA Therapy* ; Scleroderma, Localized* ; Skin

New born rat heart cells were dissociated using trypsin and/or collagenase to elucidate the dissociation efficiency of these two enzymes. And the effect of dimethyl sulfoxide during and immediately after cell dissociation was also investigated to clarify the so-called protective activity of dimethyl sulfoxide on cell performance. The results can be summarized as follows. 1. Cold trypsin 18 hours pretreatment followed by warm collagenase treatment resulted best cell viability and cell yield. 2. Single warm trypsin treatment gave the poorest result. 3. Dimethyl sulfoxide did not seem to play any protective role during or immediately after rat heart cell dissociation. It had very damaging effect on rat heart cells.
Animals ; Cell Survival ; Collagenases* ; Dimethyl Sulfoxide* ; Heart* ; Rats* ; Trypsin*

The author has observed the effects of collagenase on the relapse phenomenon and the histochemical changes during the relapse period. 50 rats were used. : 3rats as a normal group, 15rats as control groups, and 32rats as experimental groups. Rat's teeth were moved for 10days with helical spring applied, followed by injection of "collagenase in Hank's sol." to the experimental groups and the "Hank's sol." to the control group in the interdental gingiva on the 10th day, and the spring was removed on the 11th day. After injection, the experimental animals were sacrificed on the 11th, 13th, 15th, 17th, 20th, and 24th day and perpared histochemically for the Hematoylin-Eosin, Van-Gieson, and Methyl Green-pyronin staining. The results are as follows: 1. Group I (11th day): In the control group the supracrestal fibers were stretched and the metabolic rate was high. Experimental group showed that supracrestal fibers were resor, bed, disarrayed, and the metabolic rate was low. 2. Group II (13th day): In the control group, the supracrestal fibers began to vhange from the vertical direction to tooth-axis to the parallel. Experimental group showed that supracrestal fibers were completely resorbed. 3. Group IV (17th day): The control group showed almost normal structure. Form this group the metabolic rates were low. Experimental group showed the most destructive pattern. 4. Group VI (24th day): Experimental group showed almost normal structure. It follows that experimental groups were relapsed less than the control groups, and collagenase was effective in the prebention of relapse after rat's experimental tooth movement.
Animals ; Collagenases* ; Gingiva ; Rats ; Recurrence* ; Tooth Movement* ; Tooth*

The effect of orthodontic force on the collagenase and phosphatase activities of the adjacent alveolar bone was evaluated. Maxillary canines of male cats were treated orthodontically with closed coil spring so as to exert about 80g force. Sixteen cats were equally divided into one control group and seven experimental groups (12 hrs, 24 hrs, 36 hrs, 2 days, 3 days, 5 days and 7 days after orthodontic treatment). After sacrificing all animals on experimental intervals, collagenase, acid phosphatase (ACP) and alkaline phosphatase (ALP) activities were determined in the pressure and tension sides of alveolar bones. ACP activities increased in both the pressure and tension sides, but significantly increased in the pressure side continuously. ALP activities increased in the tension side at early stage (1-2 days after treatment), but changed small amount in the pressure side. Collagenase activities increased in the pressure side, especially at late stage (5-7 days after treatment). These results suggest that (1) orthodontic force increases the ACP, ALP and collagenase activities generally and (2) activities of ACP and collagenase increase in the pressure side, but that of ALP in the tension side and (3) activities of ACP and ALP increase at early stage, but that of collagenase at late stage after orthodontic treatment. Therefore it is shown that there are time differences in the formation and destruction of organic and inorganic components in the bone metabolism of alveolus with application of the orthodontic forces.
Acid Phosphatase ; Alkaline Phosphatase ; Animals ; Cats ; Collagenases ; Humans ; Male ; Metabolism

This study is to investigate the mechanism of inhibitory effect of imipramine on the calcium utilization in single cells isolated from canine detrusor. 2 mm thick smooth muscle chops were incubated in 0.12% collagenase solution at 36degreeC, and aerated with 95% O2/5% CO2, and then cell suspension was examined Acetylcholine (ACh) evoked a concentration-dependent contraction of the isolated detrusor cells in normal physiologic salt solution (PSS), and the ACh-induced contraction was significantly inhibited by imipramine. In Ca2+-free PSS, ACh-induced contraction was less than those in normal PSS and it was not affected by the pretreatment with imipramine. Ca2+-induced contraction in Ca2+-free PSS was supressed by imipramine, but addition of A 23187, a calcium ionophore, overcomed the inhibitory effect of imipramine. High potassium-depolarization (40 mM KCl) evoked cell contraction, which was inhibited by imipramine. Caffeine, a releasing agent of the stored Ca2+ from sarcoplasmic reticulum, evoked a contraction of the cells that was not blocked by the pretreatment with imipramine. These results suggest that imipramine inhibits the influx of calcium in the detrusor cells through both the receptor-operated- and voltage-gated-calcium channels, but does not affect the release of calcium from intracellular storage site.
Acetylcholine ; Caffeine ; Calcimycin ; Calcium* ; Collagenases ; Imipramine* ; Muscle, Smooth ; Sarcoplasmic Reticulum

To find out the suppressive effect of natural extract Curcuma xanthorrhiza on IL-1beta and MMP-2 derived from periodontal ligament cells through in vitro study and to confirm its effect on plaque and gingivitis through clinical study, Curcuma xanthorrhiza containing toothpaste was used and following results were produced. 1. In vitro study, type IV collagenase MMP-2 production was inhibited dose-dependently in the group treated with Curcuma xanthorrhiza compared to the control group. 2. In vitro study, the production of IL-1beta which is one of the inflammatory mediators associated with periodontitis was inhibited dose-dependently in the group treated with Curcuma xanthorrhiza. 3. On the third week, the plaque index of the groups treated with or without Curcuma xanthorrhiza containing toothpastes were both increased significantly compared to the baseline(p<0.05). 4. On the third week, the gingival index of the group treated with Curcuma xanthorrhiza containing toothpaste was not significantly different from baseline. However, the group treated without Curcuma xanthorrhiza containing toothpaste showed a significant increase of gingival index at shielded area(p<0.05). 5. The gingival index of the group without Curcuma xanthorrhiza containing toothpaste showed a significant increase in the sites without tooth brushing when compared to sites with tooth brushing(p<0.05). However, there was no significant difference for the group with Curcuma xanthorrhiza containing toothpaste in sites either with or without tooth brushing. 6. The Bleeding on probing for the group without Curcuma xanthorrhiza containing toothpaste showed no significant difference even when tooth brushing was done. However, for the group with Curcuma xanthorrhiza containing toothpaste, bleeding on probing was significantly reduced compared to baseline when tooth brushing was done(p<0.05).
Collagenases ; Curcuma* ; Gingivitis* ; Hemorrhage ; Periodontal Index ; Periodontal Ligament ; Periodontitis ; Tooth ; Toothpastes

Contraction and epithelization are two phenomena of wound healing which are retarded by steroid. Triamcinolone activates collagenase which enhanced the degradation of scar tissues. All 7 cases were treated with visual urethrotomy and injected triamcinolone into the lesion. The results were excellent :all 7 patients were asymptomatic without need of further management and their peak flow rates were within normal range. No adverse effect of intralesional injection of triamcinolone was seen.
Cicatrix ; Collagenases ; Humans ; Injections, Intralesional* ; Reference Values ; Triamcinolone* ; Urethral Stricture* ; Wound Healing

OBJECTIVETo observe the apoptosis of neural stem cells (NSCs) at differential time points after the dissociation of neurospheres by Accutase or trypsin.

METHODSThe NSCs were isolated from striatum of human fetals that suffered abortion at 12-16 weeks of pregnancy. The 3(rd)-5(th) passages of NSCs were digested by Accutase or trypsin. Only vortexing was applied, and the triturating by Pasteur pipette was avoided to attenuate the injury to the cells during the dissociation. The single cells were then stained by Annexin V/propidium iodide and Hoechst 33342. The apoptosis rates 2 and 24 hours after passaging were evaluated.

RESULTSThe trypan blue staining confirmed that immediately after the dissociation,the viability of cells digested by trypsin was (83.10 ± 6.76)%, which was significantly lower than that digested by Accutase,which was (91.65 ± 4.43)% (P<0.05). The apoptosis of the NSCs digested by Accutase was higher than that digested by trypsin at both 2 and 24 hours after passaging (P<0.01). Four days after the passaging, both the new clone formation rate and diameter of new spheres after trypsin digestion were significantly higher than those after Accutase digestion (P<0.01).

CONCLUSIONSAlthough the viability of NSCs immediately after the disassociation by trypsin is lower than that digested by Accutase, the apoptosis of NSCs subsequently caused by trypsin is lower than that caused by Accutase. Trypan blue test immediately after the disassociation can not be used as an indicator in estimating the apoptosis of NSCs during the expanding.