| There is a common clinical phenomenon: if a fractured patient has traumatic brain injury (TBI) at the same time, his fracture healing process will be accelerated and heterotopic ossification can be found near some joint without fracture. There were some clinical reports and a few experimental studies about this phenomenon, lots of hypotheses were made but the mechanism in it isn't clear so far. If traumatic brain injury makes some brain components release into the blood and there are some factors that can accelerate the fracture healing in it?Purpose:Discuss the relationship between the TBI and fracture healing, study the effect of blood plasma derived from patients with traumatic brain injury on the osteoblast, analyze the effect of the brain components on the osteoblast and the bone marrow stromal cells transforming into osteoblast. Methods: The research can be divided into 3 parts below:1. Compare the effect of tissue fluid extracted from the rat brain components (including white matter, gray matter, hypophysis, and total brain), liver, muscle, serum on rat osteoblast proliferation, differentiation, and mineralization.2. Compare the effect of tissue fluid extracted from the rat brain components (including white matter, gray matter, hypophysis, total brain), liver, muscle, serum on rat bone marrow stromal cells transforming into osteoblast and onBMSC proliferation, differentiation, and mineralization.3. Compare the effect of blood plasma derived from 3 groups (patients with TBI combined with fracture, patients with TBI only and patients with fracture only) on rat osteoblast proliferation and differentiation, and analyze the different effect on the rat osteoblast proliferation and differentiation by different period of times' blood plasma.Results:1. The effect of rat tissue fluid on rat osteoblast proliferation, differentiation, and mineralization:1) The effect on osteoblast proliferation: the gray matter group, the white matter group, the liver group and the serum group had significant difference comparing with the control group(P<0. 01), the gray matter group, the white matter group and the serum group could stimulate the osteoblast proliferation, while the liver group could inhibit the osteoblast proliferation.2) The effect on osteoblast differentiation: the total brain group, the gray matter group, the white matter group, the hypophysis group, the liver group and the serum group had significant difference comparing with the control group(P<0.01), the osteoblast ALP activity in the gray matter group, the white matter group and the serum group were higher than other groups.3) The effect on osteoblast mineralization: also the mineralization nods of the total brain group, the gray matter group, the white matter group, the muscle group were all less than the control group, but only the muscle group had the significant difference (p<0.05), showed that the muscle group could inhibit the osteoblast mineralization; the serum group had more mineralization nods than the control group, but the difference wasn' t significant.2. The effect of rat tissue fluid on rat bone marrow stromal cells transforming into osteoblast and on BMSC proliferation,differentiation, and mineralization:1) The observation of the BMSC primary culture: when the primary BMSC was cultured for 1 day, there were no significant difference in all groups, after 3 days we could see active proliferation in the gray matter group and the total brain group, after 5 days, all groups of BMSC reach confluence, but the BMSC density were higher in the gray matter group and the total brain group than in other groups, and the proliferation were more active.2) The ALP stain positive rate after primary BMSC culture: the highest ALP stain positive rate were found in the gray matter group and the total brain group, the next were the white matter group and the muscle group, and the difference was significant.3) The effect on the BMSC proliferation: all groups except the liver group could stimulate the BMSC proliferation, the gray matter group was the highest ( 204. 2% higher than the control group), then in sequence were the muscle group, the white matter group, the total brain group and the serum group, and the liver group could inhibit the BMSC proliferation.4) The effect on the BMSC differentiation: the liver group could inhibit the BMSC differentiation, while all other groups could stimulate the BMSC differentiation, their activity in descending sequence were the gray matter group, the total brain group, the white matter group, the muscle group and the serum group.5) The effect on the BMSC mineralization: the highest mineralization nods formation rate was found in the gray matter group (670.2% higher than the control group, P<0. 001), then in sequence were the total brain group, the white matter group, the muscle group and the serum group.3. The effect of blood plasma derived from 3 groups (patientswith TBI combined with fracture, patients with TBI only andpatients with fracture only) on rat osteoblast proliferationand differentiationl) The effect of all groups of blood plasma on osteoblastproliferation:a) The whole situation in all groups: the average MTT value in all groups were nearly at the first time, the MTT value in TBI combined with fracture group distinctly rose after one week and reached the peak at the second week, the MTT value in TBI only group showed downtrend after one week.b) Comparison in spite of time period, the average MTT value in TBI combined with fracture group was significant higher than in TBI only group (P<0. 01) and in fracture only group (P<0. 05).c) The MTT value difference among the time periods in each group: there was no significant difference between two time periods in the fracture only group; the average MTT value in the TBI only group rose at one week time, then descended gradually, but no significant difference among the time periods; the average MTT value in TBI combined with fracture group distinctly rose after one week and reached the peak at the second week, and the difference was significant.2) The effect of all groups of blood plasma on osteoblast differentiation:a) The whole situation in all groups: at the first time , the average ALP value in the fracture only group was highest and lowest in the TBI only group, the difference was significant; after one week, the ALP value were nearly in all groups, and the ALP value in TBI combined with fracture group rose gradually.b) Comparison in spite of time period, the average ALP value in TBI only group was significantly lower than in fracture only group (P<0.01), also lower than in TBI combined with fracture group (P<0. 05), there was no significant difference between the fracture only group and the TBI combined with fracture group.c) The ALP value difference among the time periods in each group: there was no significant difference between two time periods in the fracture only group;...
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