Bovine Insulin. Cell Culture Tested Product Code: TC431. Disclaimer: User must ensure suitability of the product(s) in their application prior to use. Products conform solely to the information contained in this and other related HiMedia™ Publications.
Bovine colostrum (BC)4 is a milk secreted during the first few days after calving and is a rich source of bioactive components such as immunoglobulins, insulin-like growth factor (I, II) (IGF I-II), transforming growth factor-β (TGF-β), platelet-derived growth factor, epidermal growth factor, tumor necrosis factor, basic fibroblast growth factor, vasoendothelial growth factor, and telomerase (1–3).
The importance of colostrum for the health of young animals has been known for a long time (1–4), but other systemic effects in adult humans fed BC orally have also been demonstrated, such as increased salivary IgA and plasma IGF-I (5), reduced symptoms of upper respiratory tract infection in humans (6), and changes in body composition of human athletes (7).
Specific growth factors have been studied in different cultured cells, such as cementoblasts (8), chondrocytes (9), fibroblasts (10), and myoblasts (11) to evaluate improved regenerative therapies for periodontal tissues, osteoarthritis, and wound healing.
The aim of this study was to evaluate the effects of a skim freeze-dried BC source (Colexan, Colostrum Technologies GmbH) rich in bioactive components (immunoglobulins, natural growth factors, and hormones), as an in vitro stimulator of canine skin fibroblast proliferation and activity. This in vitro model could be used as a model for wound repair efficiency and periodontal tissue repair for topical applications.
MATERIALS AND METHODS
Skin samples
Skin samples were taken from fresh abdominal skin obtained from surgeries in 2 different public kennels in Barcelona (Arrabassada and Granollers). The skin (n = 4) was always healthy skin from dogs between 2 and 12 y old.
Fibroblast isolation and culture
Cultures of normal canine dermal fibroblasts were established from canine skin samples taken from 4 dogs. The skin was well shaven and cleaned with 70% EtOH/Betadine before the start of cell isolation. All the fat tissue and blood vessels were removed from the skin with scissors, and then it was washed with 0.02 mol/L sodium phosphate buffer with 0.15 mol/L sodium chloride, pH adjusted to 7.4, without calcium or magnesium (PBS-/-).
Skin cells were enzymatically dispersed from skin biopsies. Briefly, washed skin was chopped into 1-mm3 fragments and incubated for 140 min in 15 mL/g skin of Dulbecco's Modified Eagle Medium (DMEM) (Gibco, Invitrogen) containing 30 mg collagenase, 18 mg hyaluronidase, 12 mg pronase, 1.5 mg DNAse (all from Sigma) supplemented with 450 mg bovine albumin and antibiotics (100 U penicillin, 100 μg streptomycin) (Gibco, Invitrogen). After digestion, cutaneous cells were washed with PBS-/-, and their viability was assessed. Cells were grown in DMEM supplemented with 5% fetal calf serum and antibiotics (all from Gibco) in 100-mm culture dishes for the initial plating and passages. Fibroblasts grew in a humidified atmosphere at 37°C with 5% CO2 for 2 d. Cells were attached, and the medium was changed twice a week until the first passage was necessary. Fibroblasts were used at passages 2–5.
Bovine colostrum source
A freeze-dried skim BC source (Colexan, Colostrum Technologies GmbH) with 85–90% total protein (85% IgG), <1% fat, 1–5% lactose, 5–7% ash, and natural growth factors and hormones (IGFI-II, 2.5–3.8 μg/g; TGF-β, 0.1–0.4 μg/g; cortisol, 170 μg/g; prolactin, 4 ng/g; estradiol, 1.98 ng/g; testosterone, 33.6 ng/g; insulin, 1.98 mU/g, and osteocalcin, 702 ng/g) was used.
MTT cell proliferation assay
Activity that stimulated cell growth was assayed with the addition of various concentrations of BC in a 96-well microplate, and the plate was incubated for 24 and 48 h at 37°C in 5% CO2. The cell proliferation in each well was estimated by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, which was first described by Mosmann in 1983 (12). This assay is based on the ability of a mitochondrial dehydrogenase enzyme from viable cells to cleave the tetrazolium rings of the pale yellow MTT and form dark blue formazan crystals, which are largely impermeable to cell membranes, thus resulting in its accumulation within healthy cells. Solubilization of the cells by the addition of a detergent (dimethyl formamide) results in the liberation of the crystals, which are solubilized. The number of surviving cells is directly proportional to the level of the formazan product created. The color can then be quantified using a simple colorimetric assay. The results can be read on a multiwell scanning spectrophotometer (ELISA reader).
Briefly, cells were plated at a density of 10,000 cells per well with the different concentrations of colostrums to be tested (0.1, 0.3, and 1 mg/mL by dry weight, respectively) or stem cell factor (SCF) (100 ng/mL, Amgen) as positive controls because it promotes cellular growth. Negative controls (without any kind of factor that stimulates the cells) were plated with the same water volumes as colostrums in the culture medium. Cells were incubated for 24 and 48 h at 37°C in 5% CO2 humidified atmosphere, and 10 μL of MTT were added to 100 μL of medium per well and incubated for 4 h until purple precipitate was visible. Fifty microliters of dimethylformamide was added and incubated at 37°C in 5% CO2 humidified atmosphere in the dark for 18 h. Finally, the absorbance was read at 570 nm. The number of surviving cells was directly proportional to the level of the formazan product created.
Statistical analysis
Results are presented as values of absorbance at 570 nm in optical density units (OD). Differences of absorbance were assessed using Student's t test; P < 0.05 was considered significant (SAS statistical package 8.2, SAS Institute).
RESULTS AND DISCUSSION
Isolated canine fibroblasts were incubated with BC at 3 different concentrations (0.1, 0.3, and 1 mg/mL) for 24 and 48 h. Canine fibroblast growth, measured by the absorbance of the different cultures (Table 1), was significantly increased in a dose-dependent manner by the 3 different concentrations after 24 h of incubation (19 to 32% related to negative control, P < 0.05). Moreover, canine fibroblast growth was still significantly stimulated in a dose-dependent manner by the 2 highest concentrations (stimulation of 12 and 32% related to negative control by 0.3 and 1 mg/mL, respectively, P < 0.05) after 48 h of incubation.
Effect on fibroblast proliferation of different concentrations of bovine colostrum added to a fibroblast culture for 24 or 48 h1
Culture time 24 h | Culture time 48 h | |||
---|---|---|---|---|
Treatment | OD | Relative activity,2% | OD | Relative activity,2% |
Control (−) | 0.343a ± 0.009 | 100 | 0.447a ± 0.007 | 100 |
Control (+) SCF,3100 ng/mL | 0.460c ± 0.020 | 134 | 0.643d ± 0.020 | 144 |
Bovine colostrum: | ||||
1 mg/mL | 0.452c ± 0.002 | 132 | 0.591c ± 0.010 | 132 |
0.3 mg/mL | 0.410b ± 0.002 | 120 | 0.501b ± 0.004 | 112 |
0.1 mg/mL | 0.408b ± 0.011 | 119 | 0.477ba ± 0.014 | 107 |
Culture time 24 h | Culture time 48 h | |||
---|---|---|---|---|
Treatment | OD | Relative activity,2% | OD | Relative activity,2% |
Control (−) | 0.343a ± 0.009 | 100 | 0.447a ± 0.007 | 100 |
Control (+) SCF,3100 ng/mL | 0.460c ± 0.020 | 134 | 0.643d ± 0.020 | 144 |
Bovine colostrum: | ||||
1 mg/mL | 0.452c ± 0.002 | 132 | 0.591c ± 0.010 | 132 |
0.3 mg/mL | 0.410b ± 0.002 | 120 | 0.501b ± 0.004 | 112 |
0.1 mg/mL | 0.408b ± 0.011 | 119 | 0.477ba ± 0.014 | 107 |
Values are means ± SEM. Cell proliferation was estimated by an MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay and is express as optical density units (OD).
The relative activity is calculated as 100 × (absorbance of a culture/absorbance of the negative control culture).
SCF: stem cell factor. Triplicates were used for each treatment. Differences of absorbance were assessed using Student's t test. Means in a row without a common superscript letter differ, P < 0.05.
Effect on fibroblast proliferation of different concentrations of bovine colostrum added to a fibroblast culture for 24 or 48 h1
Culture time 24 h | Culture time 48 h | |||
---|---|---|---|---|
Treatment | OD | Relative activity,2% | OD | Relative activity,2% |
Control (−) | 0.343a ± 0.009 | 100 | 0.447a ± 0.007 | 100 |
Control (+) SCF,3100 ng/mL | 0.460c ± 0.020 | 134 | 0.643d ± 0.020 | 144 |
Bovine colostrum: | ||||
1 mg/mL | 0.452c ± 0.002 | 132 | 0.591c ± 0.010 | 132 |
0.3 mg/mL | 0.410b ± 0.002 | 120 | 0.501b ± 0.004 | 112 |
0.1 mg/mL | 0.408b ± 0.011 | 119 | 0.477ba ± 0.014 | 107 |
Culture time 24 h | Culture time 48 h | |||
---|---|---|---|---|
Treatment | OD | Relative activity,2% | OD | Relative activity,2% |
Control (−) | 0.343a ± 0.009 | 100 | 0.447a ± 0.007 | 100 |
Control (+) SCF,3100 ng/mL | 0.460c ± 0.020 | 134 | 0.643d ± 0.020 | 144 |
Bovine colostrum: | ||||
1 mg/mL | 0.452c ± 0.002 | 132 | 0.591c ± 0.010 | 132 |
0.3 mg/mL | 0.410b ± 0.002 | 120 | 0.501b ± 0.004 | 112 |
0.1 mg/mL | 0.408b ± 0.011 | 119 | 0.477ba ± 0.014 | 107 |
Values are means ± SEM. Cell proliferation was estimated by an MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay and is express as optical density units (OD).
The relative activity is calculated as 100 × (absorbance of a culture/absorbance of the negative control culture).
SCF: stem cell factor. Triplicates were used for each treatment. Differences of absorbance were assessed using Student's t test. Means in a row without a common superscript letter differ, P < 0.05.
The results of this trial showed that freeze-dried skim BC significantly (P < 0.05) stimulated dog fibroblasts at any dose used (0.1–1 mg/mL) at 24 h culture incubation, and the effect remained significant at 48 h for the dose of 0.3 and 1 mg/mL. This activity of BC has already been shown on NIH 3T3 cells (13) and in human fibroblasts (14). Hironaka et al. (13) observed a stimulation of NIH3T cell growth only with the cream fraction of bovine colostrum and not with skim colostrum, but in our case, skimmed BC was able to stimulate cell growth at different doses.
Further investigation would be useful to identify which components of BC are efficacious in improving canine fibroblast growth.
The results of this trial showed that freeze-dried skim BC maintains activity of the bioactive substances that promote the cellular growth, suggesting potential proliferative actions by direct contact with target cells. Further studies are needed to evaluate not only local effects but any systemic effect of an oral supplementation of BC for regenerative therapies of different tissues.
LITERATURE CITED
SM
, J
.Growth factors in milk as mediators of infant development
. 1994
;:147
–.CR
, DJ
, JW
.Insulin-like growth factors (IGFs) and IGF binding proteins in mammary secretions: origins and implications in neonatal physiology
. In: MF
, B
. editors. Mechanisms regulating lactation and infant nutrients utilization.
: Wiley-Liss
; . p. 285
–.RJ
, MA
, JR
, AR
, TC
, KS
, JL
, PK
, GF
.Factors affecting insulin-like growth-I concentration in bovine colostrums
. 1991
;:2905
–.H
, P
, HS
.. Br J Nutr.
;84
(): S135
–.A
, J
, T
, T
, I
, T
, T
, S
, J
.IGF-I, IgA, and IgG responses to bovine colostrums supplementation during training
. 2002
;:732
–.GD
, JD
.Concentrated bovine colostrum protein supplementation reduces the incidence of self-reported symptoms of upper respiratory tract infection in adult males
. 2003
;:228
–.J
, MS
, D
.The effects of bovine colostrum supplementation on body composition and exercise performance in active men and women
. 2001
;:243
–.NE
, Y
, WV
, MJ
.Growth factors regulate expression of mineral associated genes in cementoblasts
. 2000
;:1591
–.E
, J
, M
, T
, K
, T
, K
.Vascular endothelial growth factor plays an important autocrine/paracrine role in the progression of osteoarthritis
. 2005
;:275
–.Z
, P
,
Q
.Influence of basic fibroblast growth factors on the migration, proliferation of osteoblast and periodontal ligament fibroblasts
. 2005
;:201
–.M
, M
, T
.Transforming growth factor-beta 1 upregulates myostatin expression in mouse C2C12 myoblasts
. 2005
;: Suppl 2
:–214
.T
.Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays
. 1983
;:55
–.T
, H
, T
.Identification and partial purification of a basic fibroblast growth factor-like growth factor derived from bovine colostrum
. 1997
;:488
–.Y
, K
, K
.Factors in bovine colostrum that enhance the migration of human fibroblasts in type I collagen gels
. 2001
;:2776
–.Abbreviations
- bovine colostrum
- IGF-II, insulin-like growth factor (I, II)
- 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
- phosphate-buffered saline buffer without calcium or magnesium
- transforming growth factor-β
Footnotes
Published in a supplement to The Journal of Nutrition. Presented as part of The WALTHAM International Nutritional Sciences Symposium: Innovations in Companion Animal Nutrition held in Washington, DC, September 15–18, 2005. This conference was supported by The WALTHAM Centre for Pet Nutrition and organized in collaboration with the University of California, Davis, and Cornell University. This publication was supported by The WALTHAM Centre for Pet Nutrition. Guest editors for this symposium were D'Ann Finley, Francis A. Kallfelz, James G. Morris, and Quinton R. Rogers. Guest editor disclosure: expenses for the editors to travel to the symposium and honoraria were paid by The WALTHAM Centre for Pet Nutrition.
Bovine colostrum (BC)4 is a milk secreted during the first few days after calving and is a rich source of bioactive components such as immunoglobulins, insulin-like growth factor (I, II) (IGF I-II), transforming growth factor-β (TGF-β), platelet-derived growth factor, epidermal growth factor, tumor necrosis factor, basic fibroblast growth factor, vasoendothelial growth factor, and telomerase (1–3).
The importance of colostrum for the health of young animals has been known for a long time (1–4), but other systemic effects in adult humans fed BC orally have also been demonstrated, such as increased salivary IgA and plasma IGF-I (5), reduced symptoms of upper respiratory tract infection in humans (6), and changes in body composition of human athletes (7).
Specific growth factors have been studied in different cultured cells, such as cementoblasts (8), chondrocytes (9), fibroblasts (10), and myoblasts (11) to evaluate improved regenerative therapies for periodontal tissues, osteoarthritis, and wound healing.
The aim of this study was to evaluate the effects of a skim freeze-dried BC source (Colexan, Colostrum Technologies GmbH) rich in bioactive components (immunoglobulins, natural growth factors, and hormones), as an in vitro stimulator of canine skin fibroblast proliferation and activity. This in vitro model could be used as a model for wound repair efficiency and periodontal tissue repair for topical applications.
MATERIALS AND METHODS
Skin samples
Skin samples were taken from fresh abdominal skin obtained from surgeries in 2 different public kennels in Barcelona (Arrabassada and Granollers). The skin (n = 4) was always healthy skin from dogs between 2 and 12 y old.
Fibroblast isolation and culture
Cultures of normal canine dermal fibroblasts were established from canine skin samples taken from 4 dogs. The skin was well shaven and cleaned with 70% EtOH/Betadine before the start of cell isolation. All the fat tissue and blood vessels were removed from the skin with scissors, and then it was washed with 0.02 mol/L sodium phosphate buffer with 0.15 mol/L sodium chloride, pH adjusted to 7.4, without calcium or magnesium (PBS-/-).
Skin cells were enzymatically dispersed from skin biopsies. Briefly, washed skin was chopped into 1-mm3 fragments and incubated for 140 min in 15 mL/g skin of Dulbecco's Modified Eagle Medium (DMEM) (Gibco, Invitrogen) containing 30 mg collagenase, 18 mg hyaluronidase, 12 mg pronase, 1.5 mg DNAse (all from Sigma) supplemented with 450 mg bovine albumin and antibiotics (100 U penicillin, 100 μg streptomycin) (Gibco, Invitrogen). After digestion, cutaneous cells were washed with PBS-/-, and their viability was assessed. Cells were grown in DMEM supplemented with 5% fetal calf serum and antibiotics (all from Gibco) in 100-mm culture dishes for the initial plating and passages. Fibroblasts grew in a humidified atmosphere at 37°C with 5% CO2 for 2 d. Cells were attached, and the medium was changed twice a week until the first passage was necessary. Fibroblasts were used at passages 2–5.
Bovine colostrum source
A freeze-dried skim BC source (Colexan, Colostrum Technologies GmbH) with 85–90% total protein (85% IgG), <1% fat, 1–5% lactose, 5–7% ash, and natural growth factors and hormones (IGFI-II, 2.5–3.8 μg/g; TGF-β, 0.1–0.4 μg/g; cortisol, 170 μg/g; prolactin, 4 ng/g; estradiol, 1.98 ng/g; testosterone, 33.6 ng/g; insulin, 1.98 mU/g, and osteocalcin, 702 ng/g) was used.
MTT cell proliferation assay
Activity that stimulated cell growth was assayed with the addition of various concentrations of BC in a 96-well microplate, and the plate was incubated for 24 and 48 h at 37°C in 5% CO2. The cell proliferation in each well was estimated by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, which was first described by Mosmann in 1983 (12). This assay is based on the ability of a mitochondrial dehydrogenase enzyme from viable cells to cleave the tetrazolium rings of the pale yellow MTT and form dark blue formazan crystals, which are largely impermeable to cell membranes, thus resulting in its accumulation within healthy cells. Solubilization of the cells by the addition of a detergent (dimethyl formamide) results in the liberation of the crystals, which are solubilized. The number of surviving cells is directly proportional to the level of the formazan product created. The color can then be quantified using a simple colorimetric assay. The results can be read on a multiwell scanning spectrophotometer (ELISA reader).
Briefly, cells were plated at a density of 10,000 cells per well with the different concentrations of colostrums to be tested (0.1, 0.3, and 1 mg/mL by dry weight, respectively) or stem cell factor (SCF) (100 ng/mL, Amgen) as positive controls because it promotes cellular growth. Negative controls (without any kind of factor that stimulates the cells) were plated with the same water volumes as colostrums in the culture medium. Cells were incubated for 24 and 48 h at 37°C in 5% CO2 humidified atmosphere, and 10 μL of MTT were added to 100 μL of medium per well and incubated for 4 h until purple precipitate was visible. Fifty microliters of dimethylformamide was added and incubated at 37°C in 5% CO2 humidified atmosphere in the dark for 18 h. Finally, the absorbance was read at 570 nm. The number of surviving cells was directly proportional to the level of the formazan product created.
Statistical analysis
Results are presented as values of absorbance at 570 nm in optical density units (OD). Differences of absorbance were assessed using Student's t test; P < 0.05 was considered significant (SAS statistical package 8.2, SAS Institute).
RESULTS AND DISCUSSION
Isolated canine fibroblasts were incubated with BC at 3 different concentrations (0.1, 0.3, and 1 mg/mL) for 24 and 48 h. Canine fibroblast growth, measured by the absorbance of the different cultures (Table 1), was significantly increased in a dose-dependent manner by the 3 different concentrations after 24 h of incubation (19 to 32% related to negative control, P < 0.05). Moreover, canine fibroblast growth was still significantly stimulated in a dose-dependent manner by the 2 highest concentrations (stimulation of 12 and 32% related to negative control by 0.3 and 1 mg/mL, respectively, P < 0.05) after 48 h of incubation.
Effect on fibroblast proliferation of different concentrations of bovine colostrum added to a fibroblast culture for 24 or 48 h1
Culture time 24 h | Culture time 48 h | |||
---|---|---|---|---|
Treatment | OD | Relative activity,2% | OD | Relative activity,2% |
Control (−) | 0.343a ± 0.009 | 100 | 0.447a ± 0.007 | 100 |
Control (+) SCF,3100 ng/mL | 0.460c ± 0.020 | 134 | 0.643d ± 0.020 | 144 |
Bovine colostrum: | ||||
1 mg/mL | 0.452c ± 0.002 | 132 | 0.591c ± 0.010 | 132 |
0.3 mg/mL | 0.410b ± 0.002 | 120 | 0.501b ± 0.004 | 112 |
0.1 mg/mL | 0.408b ± 0.011 | 119 | 0.477ba ± 0.014 | 107 |
Culture time 24 h | Culture time 48 h | |||
---|---|---|---|---|
Treatment | OD | Relative activity,2% | OD | Relative activity,2% |
Control (−) | 0.343a ± 0.009 | 100 | 0.447a ± 0.007 | 100 |
Control (+) SCF,3100 ng/mL | 0.460c ± 0.020 | 134 | 0.643d ± 0.020 | 144 |
Bovine colostrum: | ||||
1 mg/mL | 0.452c ± 0.002 | 132 | 0.591c ± 0.010 | 132 |
0.3 mg/mL | 0.410b ± 0.002 | 120 | 0.501b ± 0.004 | 112 |
0.1 mg/mL | 0.408b ± 0.011 | 119 | 0.477ba ± 0.014 | 107 |
Values are means ± SEM. Cell proliferation was estimated by an MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay and is express as optical density units (OD).
The relative activity is calculated as 100 × (absorbance of a culture/absorbance of the negative control culture).
SCF: stem cell factor. Triplicates were used for each treatment. Differences of absorbance were assessed using Student's t test. Means in a row without a common superscript letter differ, P < 0.05.
Effect on fibroblast proliferation of different concentrations of bovine colostrum added to a fibroblast culture for 24 or 48 h1
Culture time 24 h | Culture time 48 h | |||
---|---|---|---|---|
Treatment | OD | Relative activity,2% | OD | Relative activity,2% |
Control (−) | 0.343a ± 0.009 | 100 | 0.447a ± 0.007 | 100 |
Control (+) SCF,3100 ng/mL | 0.460c ± 0.020 | 134 | 0.643d ± 0.020 | 144 |
Bovine colostrum: | ||||
1 mg/mL | 0.452c ± 0.002 | 132 | 0.591c ± 0.010 | 132 |
0.3 mg/mL | 0.410b ± 0.002 | 120 | 0.501b ± 0.004 | 112 |
0.1 mg/mL | 0.408b ± 0.011 | 119 | 0.477ba ± 0.014 | 107 |
Culture time 24 h | Culture time 48 h | |||
---|---|---|---|---|
Treatment | OD | Relative activity,2% | OD | Relative activity,2% |
Control (−) | 0.343a ± 0.009 | 100 | 0.447a ± 0.007 | 100 |
Control (+) SCF,3100 ng/mL | 0.460c ± 0.020 | 134 | 0.643d ± 0.020 | 144 |
Bovine colostrum: | ||||
1 mg/mL | 0.452c ± 0.002 | 132 | 0.591c ± 0.010 | 132 |
0.3 mg/mL | 0.410b ± 0.002 | 120 | 0.501b ± 0.004 | 112 |
0.1 mg/mL | 0.408b ± 0.011 | 119 | 0.477ba ± 0.014 | 107 |
Values are means ± SEM. Cell proliferation was estimated by an MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay and is express as optical density units (OD).
The relative activity is calculated as 100 × (absorbance of a culture/absorbance of the negative control culture).
SCF: stem cell factor. Triplicates were used for each treatment. Differences of absorbance were assessed using Student's t test. Means in a row without a common superscript letter differ, P < 0.05.
The results of this trial showed that freeze-dried skim BC significantly (P < 0.05) stimulated dog fibroblasts at any dose used (0.1–1 mg/mL) at 24 h culture incubation, and the effect remained significant at 48 h for the dose of 0.3 and 1 mg/mL. This activity of BC has already been shown on NIH 3T3 cells (13) and in human fibroblasts (14). Hironaka et al. (13) observed a stimulation of NIH3T cell growth only with the cream fraction of bovine colostrum and not with skim colostrum, but in our case, skimmed BC was able to stimulate cell growth at different doses.
Further investigation would be useful to identify which components of BC are efficacious in improving canine fibroblast growth.
The results of this trial showed that freeze-dried skim BC maintains activity of the bioactive substances that promote the cellular growth, suggesting potential proliferative actions by direct contact with target cells. Further studies are needed to evaluate not only local effects but any systemic effect of an oral supplementation of BC for regenerative therapies of different tissues.
LITERATURE CITED
SM
, J
.Growth factors in milk as mediators of infant development
. 1994
;:147
–.CR
, DJ
, JW
.Insulin-like growth factors (IGFs) and IGF binding proteins in mammary secretions: origins and implications in neonatal physiology
. In: MF
, B
. editors. Mechanisms regulating lactation and infant nutrients utilization.
: Wiley-Liss
; . p. 285
–.RJ
, MA
, JR
, AR
, TC
, KS
, JL
, PK
, GF
.Factors affecting insulin-like growth-I concentration in bovine colostrums
. 1991
;:2905
–.H
, P
, HS
.. Br J Nutr.
;84
(): S135
–.A
, J
, T
, T
, I
, T
, T
, S
, J
.IGF-I, IgA, and IgG responses to bovine colostrums supplementation during training
. 2002
;:732
–.GD
, JD
.Concentrated bovine colostrum protein supplementation reduces the incidence of self-reported symptoms of upper respiratory tract infection in adult males
. 2003
;:228
–.J
, MS
, D
.The effects of bovine colostrum supplementation on body composition and exercise performance in active men and women
. 2001
;:243
–.NE
, Y
, WV
, MJ
.Growth factors regulate expression of mineral associated genes in cementoblasts
. 2000
;:1591
–.E
, J
, M
, T
, K
, T
, K
.Vascular endothelial growth factor plays an important autocrine/paracrine role in the progression of osteoarthritis
. 2005
;:275
–.Z
, P
, Q
.Influence of basic fibroblast growth factors on the migration, proliferation of osteoblast and periodontal ligament fibroblasts
. 2005
;:201
–.M
, M
, T
.Transforming growth factor-beta 1 upregulates myostatin expression in mouse C2C12 myoblasts
. 2005
;: Suppl 2
:–214
.T
.Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays
. 1983
;:55
–.T
, H
, T
.Identification and partial purification of a basic fibroblast growth factor-like growth factor derived from bovine colostrum
. 1997
;:488
–.Y
, K
, K
.Factors in bovine colostrum that enhance the migration of human fibroblasts in type I collagen gels
. 2001
;:2776
–.Abbreviations
- bovine colostrum
- IGF-II, insulin-like growth factor (I, II)
- 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
- phosphate-buffered saline buffer without calcium or magnesium
- transforming growth factor-β
Footnotes
Published in a supplement to The Journal of Nutrition. Presented as part of The WALTHAM International Nutritional Sciences Symposium: Innovations in Companion Animal Nutrition held in Washington, DC, September 15–18, 2005. This conference was supported by The WALTHAM Centre for Pet Nutrition and organized in collaboration with the University of California, Davis, and Cornell University. This publication was supported by The WALTHAM Centre for Pet Nutrition. Guest editors for this symposium were D'Ann Finley, Francis A. Kallfelz, James G. Morris, and Quinton R. Rogers. Guest editor disclosure: expenses for the editors to travel to the symposium and honoraria were paid by The WALTHAM Centre for Pet Nutrition.