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. 2023 May 1;34(8):1335–1353. doi: 10.1007/s00198-023-06709-7

Table 2.

Characteristics of the included systematic reviews

Author, year Study type, study period Study population Exposition Protein intake Outcome Effect estimates Heterogeneity estimators NutriGrade rating AMSTAR 2 rating
Blair 2020

- SR without MA of RCTs

- Published before 09/2018

- Study duration: 6–18 mos

- Both sexes

- Healthy people

- Mean age: 50–70.5 yrs

Whey protein Supplements: 40 mg/d–40 g/d NA High
2 RCTs n = 148 BMD 2 out of 2 RCTs reported no changes to BMD Low: 4.5
2 RCTs n = 148 Osteocalcin 2 out of 2 RCTs reported unchanged osteocalcin Low: 4.5
Darling 2019

- SR with MA of RCTs and observational studies

- Published between 01/1976 and 02/2016

- Study duration: 38 d–2 yrs

- Follow-up: 1–22 yrs

- Both sexes

- Healthy people

- Aged 20–82 yrs

Supplements in RCTs: 40 mg/d milk-based protein or 20.4–45 g/d protein or 18–40 g/d soy protein

Cohort studies: total protein (mean): ≥ 46 g/d animal protein (mean): 17.6–29.0 g/d or 0.8–1.2 g/kg BW/d plant protein (mean): 12 g/d or 1.2 g/kg BW/d

Low
4 cohort studies n = 157,046 Total protein Low trauma fractures

RR (95% CI), fixed effect 1.04 (0.93, 1.16)

P = NP

I2 = 32%

tau2 = 0.026

P = 0.22

Very low: 2.0
4 cohort studies n = 193,954 Animal protein Low trauma fractures

RR (95% CI), random effect 0.98 (0.76, 1.27)

P = 0.87

I2 = 46%

tau2 = 0.031

P = 0.13

Very low: 2.0
3 cohort studies n = 154,167 Plant protein Low trauma fractures

RR (95% CI), fixed effect 0.97 (0.87, 1.09)

P = 0.61

I2 = 15%

tau2 = 0.007

P = 0.31

Very low: 2.0
2 RCTs n = 255 High vs low intake of total protein Lumbar spine BMD

MD (95% CI), fixed effect 0.04 g/cm2 (0.00, 0.08)

P = 0.24

I2 = 0%

tau2 = 0

P = 0.47

Very low: 2.0
3 RCTs n = 435 High vs low intake of total protein Femoral neck BMD

MD (95% CI), random effect 0.01 g/cm2 (− 0.03, 0.05)

P = 0.07

I2 = 68%

tau2 = 0.001

P = 0.04

Very low: 2.0
3 cohort studies n = 11,112 Total protein Fracture risk

HR (95% CI), fixed effect 0.83 (0.67, 1.01)

P = 0.59

I2 = 35%

tau2 = 0.048

P = 0.19

Very low: 2.0
3 RCTs n = 115 Milk basic protein Lumbar spine BMD

MD (95% CI), fixed effect 0.02 g/cm2 (0.00, 0.04)

P = 0.08

I2 = 0%

tau2 = 0

P = 0.87

Very low: 2.0
NP NP Soy protein Lumbar spine BMD

MD (95% CI), random effect − 0.01 g/cm2 (− 0.07, 0.06)

P = 0.82

I2 = 51%

P = 0.13

Very low: (2.0)
NP NP Soy protein Femoral neck BMD

MD (95% CI), random effect 0.01 g/cm2 (− 0.06, 0.07)

P = 0.87

I2 = 74%

P = 0.05

Very low: 2.0
6 RCTs n = 128 Soy protein BAP

MD (95% CI), random effect − 1.75 ng/ml (− 10.50, 7.01)

P = 0.70

I2 = 91%

P = 0.00

Very low: 2.7
Groenendijk 2019

- SR with MA of cohort studies

- Published before 11/2018

- Follow-up: 1–32 yrs

- Both sexes

- Healthy people

- Aged ≥ 50 yrs

High vs low protein

Cohort studies (mean intake)

≥ 0.93 g/kg BW/d vs < 0.95 g/kg BW/d or 19.5–29.1 E% vs 9.6–13.3 E% or 82.7 g/d vs 46.5 g/d

NA High
2 cohort studies n = 1406 Total body BMC One cohort study found a significant higher total body BMC, whereas one cohort study found no association Very low: 2.0
4 cohort studies n = 145,558 Total body BMD 3 out of 4 cohort studies found no association Low: 4.0
5 cohort studies n = 153,841 Total hip BMD 3 out of 5 cohort studies found no association Low: 4.0
6 cohort studies n = 4887 Femoral neck BMD Half of the cohort studies showed significantly higher femoral neck BMD and the other half showed no association Low: 3.0
4 cohort studies n = 1593 Lumbar spine BMD 2 cohort studies found no association, whereas one cohort study found significantly higher and one cohort study found significantly lower lumbar spine BMD Very low: 2.0
2 cohort studies n = 434 Osteocalcin Both cohort studies showed no association Low: 3.0
2 cohort studies n = 434 NTX Both cohort studies showed no association Low: 3.0
MA of 4 cohort studies n = 144,580 Hip fracture

Weighted HR (95% CI), fixed effect 0.89 (0.84, 0.94)

P < 0.001

I2 = 0%

P = 0.61

Very low: 2.5
Wright 2019

- SR with MA of RCTs

- Published before 01/2019

- Study duration: 3 mos–2 yrs

- Both sexes

- People, which actively losing weight (BMI 31.4 ± 2.5 kg/m2)

- Aged 49 ± 10 yrs

High protein diet: dietary protein ≥ 25 En% and/or ≥ 1 g/kg BW/d 1.01–1.69 g/kg BW/d vs 0.67–0.97 g/kg BW/d MD (95% CI), random effect Low
8 RCTs n = 488 Total body BMD

0.006 g/cm2 (0, 0.012)

P = 0.0163

I2 = 69%

tau2 = 0

P = 0.001

Low: 5.65
2 RCTs n = 184 (only postmenopausal women) Total hip BMD

0 g/cm2 (− 0.026, 0.026)

P = 0.497

I2 = 0%

tau2 = 0

P = 1.00

Low: 5.5
3 RCTs n = 272 Femoral neck BMD

0 g/cm2 (− 0.016, 0.016)

P = 0.50

I2 = 0%

tau2 = 0

P = 1.00

Low: 5.75
4 RCTs n = 322 Lumbar spine BMD

0.017 g/cm2 (0.001, 0.033)

P = 0.0187

I2 = 0%

tau2 = 0

P = 0.72

low: 5.25
7 RCTs n = 408 Total body BMC

4.28 g (− 11.49, 20.04)

P = 0.393

I2 = 0%

tau2 = 0

P = 0.94

Low: 5.4
2 RCTs n = 135 Femoral neck BMC

− 0.05 g (− 0.16, 0.05)

P = 0.848

I2 = 0%

tau2 = 0

P = 0.78

Low: 5.62
2 RCTs n = 135 Lumbar spine BMC

0.40 g (− 1.19, 2.00)

P = 0.310

I2 = 0%

tau2 = 0

P = 0.81

Low: 5.25
Shams-White 2018

- SR without MA of RCTs

- Published before 11/2016

- Study duration: 6–24 mos

- Peri- or postmenopausal women

- Generally healthy people

- Mean age: 50.6–73.1 yrs

Supplements: 18–40 g/d Low
2 RCTs n = 393 Isoflavone-poor soy protein vs animal protein Lumbar spine BMD Both RCTs found no significant differences in the net changes in lumbar spine BMD NA Moderate: 5.5
2 RCTs n = 146 Isoflavone-poor soy protein vs animal protein Femoral neck BMD Both RCTs found no significant differences in the net changes in femoral neck BMD NA Moderate: 5.5
2 RCTs n = 146 Isoflavone-poor soy protein vs animal protein Total body BMD Both RCTs found no significant differences in the net changes in total body BMD NA Moderate: 5.5
2 RCTs n = 91 Isoflavone-poor soy protein vs animal protein BAP Both RCTs found no significant differences in the net changes in BAP NA Low: 5.25
2 RCTs n = 133 Isoflavone-poor soy protein vs animal protein NTX Both RCTs found no significant differences in the net changes in NTX NA Low: 5.25
Shams-White 2017

- SR with MA of RCTs

- Published between 1946 and 10/2016

- Study duration: 6–24 mos

- Both sexes

- Healthy people

- Aged ≥ 18 yrs

Higher protein vs lower protein 10–45 g/d vs. 0–2.1 g/d or 25–30 E% vs. 15–18 E% or 1.4 g/kg BW/d vs. 0.8 g/kg BW/d Pooled net change (95% CI), random effect Low
5 RCTs n = 989 Lumbar spine BMD

0.52% (0.06, 0.97)

P = NP

I2 = 0%

P = 0.58

Low: 5.0
7 RCTs n = 1205 Total hip BMD

0.30% (− 0.02, 0.62)

P = NP

I2 = 0%

P = 0.54

Low: 4.4
6 RCTs n = 1169 Femoral neck BMD

− 0.14% (− 0.60, 0.32)

P = NP

I2 = 0%

P = 0.95

Low: 4.4
8 RCTs n = 494 Osteocalcin

0.06 ng/ml (− 0.49, 0.60)

P = NP

I2 = 27%

P = 0.21

Low: 4.4
5 RCTs n = 370 CTX

47.72 ng/L (− 27.34, 122.78)

P = NP

I2 = 61.3%

P = 0.04

Low: 4.0
Wu 2015

- SR with MA of cohort studies

- Published before 07/2014 and 10/2016

- Follow-up: NP

- Both sexes

- Aged 18–89 yrs

Total protein: 50.11–> 98 g/d vs < 40.75–< 68 g/d or 20% increased (g/d) vs normal

Animal protein: > 20.6–> 80 g/d vs < 13.6–< 51 g/d or 1 g/kg BW/d increased vs normal

Plant protein: > 13.27–> 26.2 g/d vs < 4.98–18 g/d or 1 g/kg BW/d increased vs normal

Adjusted RR (95% CI), random effect High
3 cohort studies n = 215,493 Total protein All fractures

0.99 (0.97, 1.02)

P = NP

I2 = 0%

P = 0.57

Very low: 3.5
2 cohort studies n = 36,342 Animal protein All fractures

0.79 (0.32, 1.96)

P = NP

I2 = 70%

P = 0.07

Very low: 3.5
3 cohort studies n = 60,745 Plant protein All fractures

0.77 (0.52, 1.12)

P = NP

I2 = 86%

P = 0.001

Very low: 3.5
6 cohort studies n = 270,011 Total protein Hip fractures

0.89 (0.82, 0.97)

P = NP

I2 = 0%

P = 0.44

Low: 4.0
4 cohort studies n = 161,393 Animal protein Hip fractures

1.04 (0.70, 1.54)

P = NP

I2 = 52%

P = 0.08

Very low: 2.5
3 cohort studies n = 121,606 Plant protein Hip fractures

1.00 (0.53, 1.91)

P = NP

I2 = 57%

P = 0.01

Very low: 2.5
2 cohort studies n = 230,480 Total protein Limb fractures

1.05 (0.81, 1.37)

P = NP

I2 = 90%

P = 0.01

Very low: 2.5
2 cohort studies n = 87,765 Plant protein Limb fractures

0.94 (0.40, 2.22)

P = NP

I2 = 86%

P = 0.001

Very low: 2.5
Pedersen 2014

- SR without MA of RCTs and cohort studies

- Published between 01/2000 and 01/2012

- Study duration: 63 d–3 yrs

- Follow-up: 1–7 yrs

- Both sexes

- Healthy people

- Aged 55–92 yrs

High vs low protein

Cohorts: 82.7–> 87 g/d vs 46.5–< 66 g/d

1.2–> 1.6 g/kg BW/d vs 0.84–1.1 g/kg BW/d

RCTs: 1.2 g/kg BW/d vs 1.1 g/kg BW/d or 20–24 E% vs 14–16 E%

NA High
1 RCT, 3 cohort studies n = 2543 Total protein BMD “The evidence is assessed as suggestive regarding a positive association between protein intake and BMD.” Moderate: 4.5
1 RCT, 3 cohort studies n = 1621 Total protein Bone loss 3 out of 4 studies found no statistically significant associations between total protein and bone loss Low: 3.0
1 RCT, 2 cohort studies n = 1529 Animal protein 2 out of 3 studies found no statistically significant associations between total protein and bone loss Low: 3.0
Plant protein None of the studies found significant associations between total protein and bone loss Low: 3.0
1 cohort study n = 946 Total protein Fracture risk This cohort study found an inverse association NA
1 cohort study n = 1035 Animal protein Fracture risk This cohort study found a positive association NA
1 cohort study n = 807 Total, animal and plant protein Falls This cohort study found no significant associations between falls and total, animal and plant protein NA
Pedersen 2013

- SR without MA of cohort studies

- Published between 01/2000 and 12/2011

- Follow-up: 2–15 yrs

- Both sexes

- Healthy and/or people with prehypertension or stage 1 hypertension

- Aged 14–69 yrs

- n = NP for all studies

High vs low protein 17.4-30.8 En% vs 5.6–19.9 En% or > 82.7 g/d vs 46.5–65 g/d or > 1.2 g/kg BW/d vs 0.84–1.1 g/kg BW/d NA Moderate
3 cohort studies n = 2210 Total protein Bone loss 2 out of 3 cohort studies found no significant associations between total protein and bone loss Low: 3.0
3 cohort studies n = 40,908 Total protein Fracture risk None of the cohort studies found significant associations Moderate: 5.0
1 cohort study n = 1,035 Animal protein Fracture risk This cohort study found a positive association NA
1 cohort study n = 1,035 Plant protein Fracture risk This cohort study found no significant associations NA
Santesso 2012

- SR with MA of RCTs

- Published before 08/2011

- Study duration: 63–180 d

- Both sexes

- Healthy and/or people with overweight, obesity hyperlipidemia, hypertension or MS

- Aged 26–54 yrs

High protein diet vs low protein diet Median: 27 En% (range: 16–45 En%) vs. 18 En% (range: 5–23 En%) MD (95% CI), random effect High
3 RCTs n = 208 Total body BMD

0.00 g/cm2 (− 0.02, 0.03)

P = 0.82

I2 = 0%

tau2 = 0.00

P = 0.74

Very low: 3.0
2 RCTs n = 180 Lumbar spine BMD

0.60 g/cm2 (− 0.34, 1.54)

P = 0.21

I2 = 67%

tau2 = 0.45

P = 0.05

Very low: 3.0
1 RCT n = 130 Hip BMD

0.61 g/cm2 (− 0.13, 1.34)

P = 0.11

I2 = 0%

tau2 = 0.00

P = 0.79

Very low: 3.0
Darling 2009

- SR with MA of RCTs and cohort studies

- Published between 1974 and 07/2008

- Study duration: 38 d–6 mos

- Follow-up: 1–22 yrs

- Both sexes

- Healthy people

- Aged 35–91 yrs

Higher vs lower protein

Supplements in RCTs: 40 mg/d milk-based protein or 20.4 g/d protein

Cohort studies: median: 79.6 g/d

Moderate
3 cohort studies n = 120,849 Total protein Hip fractures

RR (95% CI), random effect 0.75 (0.47, 1.20)

P = 0.23

I2 = 20.4%

P = 0.28

Low: 4.5
3 cohort studies n = 157,737 Animal protein Hip fractures

RR (95% CI), random effect 0.83 (0.54, 1.30)

P = 0.42

I2 = 48.3%

P = 0.14

Very low: 3.5
2 cohort studies n = 117,950 Plant protein Hip fractures

RR (95% CI), random effect 1.21 (0.82, 1.79)

P = 0.34

I2 = 2.0%

P = 0.31

Very low: 3.5
3 RCTs n = 110 Protein supplementation Lumbar spine BMD

MD (95% CI), fixed effect 0.02 g/cm2 (0.00, 0.04)

P = 0.04

I2 = 0%

P = 0.62

Very low: 3.5

Abbreviations: AMSTAR 2 A Measurement Tool to Assess Systematic Reviews, BAP bone-specific alkaline phosphatase, BMC bone mineral content, BMD bone mineral density, BW body weight, CI confidence interval, CTX C-terminal telopeptide, d day(s), En% energy percentage, HR hazard ratio, MA meta-analysis, MD mean difference, mo month, MS metabolic syndrome, NA not applicable, NP not provided, NTX N-terminal telopeptide, RCT randomized controlled trial, RR relative risk, SR systematic review, WMD weighted mean difference, wk week, yr year