J Food Bioact

Journal of Food Bioactives, ISSN 2637-8752 print, 2637-8779 online

Journal website www.isnff-jfb.com

Review

Volume 3, Number , September 2018, pages 101-110

Gut microbiota, probiotics, prebiotics and bone health: a review

Figure 1.
The Sketch of the regulators of gut microbiome and the relationship between gut microbiome and bone metabolism.
Both host and environment affects the individual gut microbiota and further regulate bone metabolism via immune system, endocrine system and calcium absorption. (Modified according to Xu et al., 2017).

**Table 1.** Current study on gut microbiota and bone health
Research model	Treatment	Outcomes	Reference
Human study
Human Predispose to bacterial overgrowth	N/A	Low BMD in lumbar and femoral High bone loss at lumbar and femoral Small intestine bacterial overgrowth is a cofactor in the development of metabolic bone disease	Di Stefano et al., 2001
Human Predispose to bacterial overgrowth	N/A	Low BMD in lumbar and femoral 43% of the patients had had fractures	Stotzer et al., 2003
Human Small bowel bacterial overgrowth (SBBO)	N/A	Significant difference in femoral BMD between healthy control and SBBO-negative group and SBBO-positive group No significant difference in femoral BMD between SBBO-positive group and SBBO-negative group	Mitsui et al., 2005
Germ-free Animal Study and Antibiotics Intervention Study
C57B16/J mice Female germ-free	N/A	Increased bone mass in germ-free mice associated with reduced number of osteoclasts Colonization of germ-free mice with normal gut microbiota normalizes bone mass	Sjögren et al., 2012
C57BL/6J mice Male and female	Exposed to low-dose penicillin post-weaning (LDP-w)	LDP-w male mice showed decreased bone mineral content LDP-w female mice showed elevated bone mineral density	Cox et al., 2014
C57BL/6J mice Male and female	Pulsed antibiotic treatemetn (PAT) with 1) amoxicillin; 2) tylosin; 3) sequential courses of tylosin, amoxicillin and tylosin	PAT accelerated bone growth and caused progressive changes in gut microbiome diversity, population structure and metagenomic content	Nobel et al., 2015
BALB/c mice Male germ-free	Lactobacillus. Plantarum^WJL and L. plantarum^NIZO2877 were gavage to 8-week-old mice to develop Lactobacillus-monocolonized mice	Reduced femur length, cortical thickness, cortical bone fraction, and the trabecular fraction of the femur in germ-free mice Lactobacilli promoted juvenile growth	Schwarzer et al., 2016
C57BL/6 mice Male germ-free	Microbiota from 6-month/8-month healthy or undernourished infants were gavage to germ-free mice	Higher femoral cortical ratios of bone volume to tissue volume (BV/TV) and volumetric bone mineral density (vBMD) in recipients of undernourished donor gut microbiota Mice colonized with 6-month old donor microbiota had higher BMD, BV/TV, trabecular connectivity and number and lower trabecular spacing	Blanton et al., 2016
CB6F1 mice Male germ-free	Microbiota from 3-month-old male CB6F1 SPF mice were gavage to germ-free mice	Increased both formation and resorption in microbiota colonized group Acutely reduced bone loss after colonization Increased bone formation, growth plate activity and insulin-like growth factor 1(IGF-1) showed in long-term colonization	Yan et al., 2016
C57BL6/J mice Female germ-free	Microbiota from 10-week-old C57BL6 mice were gavage to germ-free mice GnRH agonists Lupron Depot was administered to induce sex steroid deficiency Sex steroid deficiency mice treated with Lactobacillus rhamnosus GG, commercial probiotic supplement VSL#3, Escherichia coli and a mutant L. rhamnosus GG	Germ-free mice failed to increase sex steroid deficiency-induced bone resorption and trabecular bone loss Probiotic treatment reduced bone marrow inflammation and protected against bone loss nonprobiotic (E. coli and mutant L. rhamnosus GG) treatment showed no protective effect	Li et al., 2016
Swiss Wester mice C57BL/6 mice Germ-free	Distinct microbiotas that originated from human or mice were gavage to germ-free mice	No significantly change between germ-free group and colonized group	Quach et al., 2018

Table 1. Current study on gut microbiota and bone health

Research model

Treatment

Outcomes

Reference

Human study

Human
Predispose to bacterial overgrowth

N/A

Low BMD in lumbar and femoral
High bone loss at lumbar and femoral
Small intestine bacterial overgrowth is a cofactor in the development of metabolic bone disease

Di Stefano et al., 2001

Human
Predispose to bacterial overgrowth

N/A

Low BMD in lumbar and femoral
43% of the patients had had fractures

Stotzer et al., 2003

Human
Small bowel bacterial overgrowth (SBBO)

N/A

Significant difference in femoral BMD between healthy control and SBBO-negative group and SBBO-positive group
No significant difference in femoral BMD between SBBO-positive group and SBBO-negative group

Mitsui et al., 2005

Germ-free Animal Study and Antibiotics Intervention Study

C57B16/J mice
Female germ-free

N/A

Increased bone mass in germ-free mice associated with reduced number of osteoclasts
Colonization of germ-free mice with normal gut microbiota normalizes bone mass

Sjögren et al., 2012

C57BL/6J mice
Male and female

Exposed to low-dose penicillin post-weaning (LDP-w)

LDP-w male mice showed decreased bone mineral content
LDP-w female mice showed elevated bone mineral density

Cox et al., 2014

C57BL/6J mice
Male and female

Pulsed antibiotic treatemetn (PAT) with 1) amoxicillin; 2) tylosin; 3) sequential courses of tylosin, amoxicillin and tylosin

PAT accelerated bone growth and caused progressive changes in gut microbiome diversity, population structure and metagenomic content

Nobel et al., 2015

BALB/c mice
Male germ-free

Lactobacillus. Plantarum^WJL and L. plantarum^NIZO2877 were gavage to 8-week-old mice to develop Lactobacillus-monocolonized mice

Reduced femur length, cortical thickness, cortical bone fraction, and the trabecular fraction of the femur in germ-free mice
Lactobacilli promoted juvenile growth

Schwarzer et al., 2016

C57BL/6 mice
Male germ-free

Microbiota from 6-month/8-month healthy or undernourished infants were gavage to germ-free mice

Higher femoral cortical ratios of bone volume to tissue volume (BV/TV) and volumetric bone mineral density (vBMD) in recipients of undernourished donor gut microbiota
Mice colonized with 6-month old donor microbiota had higher BMD, BV/TV, trabecular connectivity and number and lower trabecular spacing

Blanton et al., 2016

CB6F1 mice
Male germ-free

Microbiota from 3-month-old male CB6F1 SPF mice were gavage to germ-free mice

Increased both formation and resorption in microbiota colonized group
Acutely reduced bone loss after colonization
Increased bone formation, growth plate activity and insulin-like growth factor 1(IGF-1) showed in long-term colonization

Yan et al., 2016

C57BL6/J mice
Female germ-free

Microbiota from 10-week-old C57BL6 mice were gavage to germ-free mice
GnRH agonists Lupron Depot was administered to induce sex steroid deficiency
Sex steroid deficiency mice treated with Lactobacillus rhamnosus GG, commercial probiotic supplement VSL#3, Escherichia coli and a mutant L. rhamnosus GG

Germ-free mice failed to increase sex steroid deficiency-induced bone resorption and trabecular bone loss
Probiotic treatment reduced bone marrow inflammation and protected against bone loss
nonprobiotic (E. coli and mutant L. rhamnosus GG) treatment showed no protective effect

Li et al., 2016

Swiss Wester mice
C57BL/6 mice
Germ-free

Distinct microbiotas that originated from human or mice were gavage to germ-free mice

No significantly change between germ-free group and colonized group

Quach et al., 2018

**Table 2.** current studies on probiotics/prebiotics and bone health
Research model	Treatment	Outcomes	Reference
Probiotics
C57B1/6 mice Male and female	Treated with Lactobacillus reuteri ATCC PTA 6475	L. reuteri 6475 treatment increased male trabecular bone parameters in the distal femur metaphysieal regions and the lumbar vertebrae in male mice L. reuteri 6475 treatment had no effect on bone parameters in female mice	McCabe et al., 2013
Balb/c mice Female OVX	Treated with Lactobacillus reuteri ATCC PTA 6475	L. reuteri treatment protected OVX mice from bone loss L. reuteri treatment decreased osteoclastogenesis	Britton et al., 2014
B57BL/6N mice Female OVX	Treated with 1) Lactobacillus paracasei DSM13423; 2) a mixture of L. paracasei DSM13434, L. plantarum DSM 15312 and DSM 15313	L. paracasei DSM13423 and the mixture treatment protected from OVX-induced cortical bone loss and bone resorption	Ohlsson et al., 2014
Balb/c mice Female with mild inflammatory state induced by dorsal surgical incision (DSI)	Orally supplemented with L. reuteri PTA 6457	L. eruteri 6457 treatment increased bone density in females but only in those that underwent DSI	Collins et al., 2016
BALB/c mice Male germ-free	Lactobacillus. Plantarum^WJL and L. plantarum^NIZO2877 were gavage to 8-week-old mice to develop Lactobacillus-monocolonized mice	Reduced femur length, cortical thickness, cortical bone fraction, and the trabecular fraction of the femur in germ-free mice Lactobacilli promoted juvenile growth	Schwarzer et al., 2016
C57BL6/J mice Female germ-free	Sex steroid deficiency mice treated with Lactobacillus rhamnosus GG, commercial probiotic supplement VSL#3, Escherichia coli and a mutant L. rhamnosus GG	Probiotic treatment reduced bone marrow inflammation and protected against bone loss nonprobiotic (E. coli and mutant L. rhamnosus GG) treatment showed no protective effect	Li et al., 2016
Prebiotics
Wistar rat Female ovariectomized (OVX)	Treated with galactooligosaccharides (GOS)	GOS treatment absorbed calcium more efficiently GOS showed higher bone (femur and tibia) ash weight and tibia calcium context of OVX rats	Chonan et al., 1995
Wistar rat Male rat	Treated with inulin and resistant starch	Inulin and resistant starch increased the intestinal absorption and balance of Ca and Mg, without altering the plasma level of Ca and Mg	Younes et al., 2001
Sprague Dawley rat Female OVX	Treated with inulin and fructooligosaccharides (FOS)	Inulin and FOS treatment increased bone mineral density, breaking strength, and total calcium Inulin and FOS treatment inhibited the bone resorption	Zafar et al., 2004
Sprague Dawley rat Male rat	Treated with inulin-type fructans (inulin, oligofructose, fructooligosaccharides)	Inulin-type fructans increased mineral absorption, retention and accumulation in bone in the case of Ca, Mg and Zn	Raschka and Daniel, 2005
Sprague Dawley rats Female OVX	Treated with vitamin D-deficient diet with or without difructose anhydride III (DFAIII)	Vitamin D deficiency decreased Ca absorption and bone mineralization DFAIII treatment restored the reduction of Ca absorption and femoral Ca	Mitamura and Hara, 2006
BALB/c mice Female	Treated with antibiotics and short chain fatty acid (SCFA)	Antibiotic treatment decreased serum IGF-1 and inhibited bone formation Antibiotic treatment group supplemented with SCFA restored IGF-1 and bone mass	Yan et al., 2016
Human Postmenopausal women	Supplemented with inulin	Inulin treatment improved mineral absorption and impacted bone turnover	Holloway et al., 2007
Human Postmenopausal women	Treated with 1) milk fermented with Lactobacillus helveticus LBK-16H bacteria, supplemented with 14.5mg/100g of IPP and VPP; 2) normal sour milk fermented with a Lactococcus sp. Mixed culture; 3) orange juice with 14.5mg/100g of IPP and VPP; 4) orange juice with calcium lactate gluconate	L. helveticus fermented milk treatment reduced serum PTH and increased serum calcium	Narva et al., 2004b

Table 2. current studies on probiotics/prebiotics and bone health

Research model

Treatment

Outcomes

Reference

Probiotics

C57B1/6 mice
Male and female

Treated with Lactobacillus reuteri ATCC PTA 6475

L. reuteri 6475 treatment increased male trabecular bone parameters in the distal femur metaphysieal regions and the lumbar vertebrae in male mice
L. reuteri 6475 treatment had no effect on bone parameters in female mice

McCabe et al., 2013

Balb/c mice
Female OVX

Treated with Lactobacillus reuteri ATCC PTA 6475

L. reuteri treatment protected OVX mice from bone loss
L. reuteri treatment decreased osteoclastogenesis

Britton et al., 2014

B57BL/6N mice
Female OVX

Treated with 1) Lactobacillus paracasei DSM13423; 2) a mixture of L. paracasei DSM13434, L. plantarum DSM 15312 and DSM 15313

L. paracasei DSM13423 and the mixture treatment protected from OVX-induced cortical bone loss and bone resorption

Ohlsson et al., 2014

Balb/c mice
Female with mild inflammatory state induced by dorsal surgical incision (DSI)

Orally supplemented with L. reuteri PTA 6457

L. eruteri 6457 treatment increased bone density in females but only in those that underwent DSI

Collins et al., 2016

BALB/c mice
Male germ-free

Lactobacillus. Plantarum^WJL and L. plantarum^NIZO2877 were gavage to 8-week-old mice to develop Lactobacillus-monocolonized mice

Reduced femur length, cortical thickness, cortical bone fraction, and the trabecular fraction of the femur in germ-free mice
Lactobacilli promoted juvenile growth

Schwarzer et al., 2016

C57BL6/J mice
Female germ-free

Sex steroid deficiency mice treated with Lactobacillus rhamnosus GG, commercial probiotic supplement VSL#3, Escherichia coli and a mutant L. rhamnosus GG

Probiotic treatment reduced bone marrow inflammation and protected against bone loss
nonprobiotic (E. coli and mutant L. rhamnosus GG) treatment showed no protective effect

Li et al., 2016

Prebiotics

Wistar rat
Female ovariectomized (OVX)

Treated with galactooligosaccharides (GOS)

GOS treatment absorbed calcium more efficiently
GOS showed higher bone (femur and tibia) ash weight and tibia calcium context of OVX rats

Chonan et al., 1995

Wistar rat
Male rat

Treated with inulin and resistant starch

Inulin and resistant starch increased the intestinal absorption and balance of Ca and Mg, without altering the plasma level of Ca and Mg

Younes et al., 2001

Sprague Dawley rat
Female OVX

Treated with inulin and fructooligosaccharides (FOS)

Inulin and FOS treatment increased bone mineral density, breaking strength, and total calcium
Inulin and FOS treatment inhibited the bone resorption

Zafar et al., 2004

Sprague Dawley rat
Male rat

Treated with inulin-type fructans (inulin, oligofructose, fructooligosaccharides)

Inulin-type fructans increased mineral absorption, retention and accumulation in bone in the case of Ca, Mg and Zn

Raschka and Daniel, 2005

Sprague Dawley rats
Female OVX

Treated with vitamin D-deficient diet with or without difructose anhydride III (DFAIII)

Vitamin D deficiency decreased Ca absorption and bone mineralization
DFAIII treatment restored the reduction of Ca absorption and femoral Ca

Mitamura and Hara, 2006

BALB/c mice
Female

Treated with antibiotics and short chain fatty acid (SCFA)

Antibiotic treatment decreased serum IGF-1 and inhibited bone formation
Antibiotic treatment group supplemented with SCFA restored IGF-1 and bone mass

Yan et al., 2016

Human
Postmenopausal women

Supplemented with inulin

Inulin treatment improved mineral absorption and impacted bone turnover

Holloway et al., 2007

Human
Postmenopausal women

Treated with 1) milk fermented with Lactobacillus helveticus LBK-16H bacteria, supplemented with 14.5mg/100g of IPP and VPP; 2) normal sour milk fermented with a Lactococcus sp. Mixed culture; 3) orange juice with 14.5mg/100g of IPP and VPP; 4) orange juice with calcium lactate gluconate

L. helveticus fermented milk treatment reduced serum PTH and increased serum calcium

Narva et al., 2004b