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Subgroup analysis of patients with previously performed abdominal surgery n = 381 revealed significantly higher incisional herniation rates when “advanced” diabetic nephropathy was pre

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DOI 10.1007/s11255-016-1229-8

NEPHROLOGY - ORIGINAL PAPER

Effects of various stages of nephropathy on wound healing

in patients with diabetes: an observational cohort study

encompassing 731 diabetics

Paula Loewe 1 · Ioannis Stefanidis 1 · Peter R Mertens 1 · Christos Chatzikyrkou 1

Received: 1 October 2015 / Accepted: 25 January 2016

© Springer Science+Business Media Dordrecht 2016

herniation, aortic aneurysms and varicose veins, did not occur more frequently than in patients without nephropa-thy In diabetics with nephropathy, umbilical herniation

(3 vs 8.2 %, p = 0.04) and disc herniation rates (5.7 vs 16.1 %, p = 0.002) were significantly lower Subgroup

analysis of patients with previously performed abdominal

surgery (n = 381) revealed significantly higher incisional

herniation rates when “advanced” diabetic nephropathy was present (16 % compared to 5.7 % without nephropathy,

p = 0.016)

Conclusion Our findings support the hypothesis that inci-sional hernia formation and diabetic nephropathy are posi-tively correlated Conversely, umbilical and disc herniation pathomechanisms are distinct, as these negatively correlate with the presence of advanced diabetic nephropathy

Keywords Diabetic nephropathy · Chronic kidney

disease · Incisional hernia · Connective tissue disease · Risk factor

Introduction

Diabetic nephropathy is a microvascular complication characterized by albuminuria, arterial hypertension and a progressive decline in glomerular filtration rate [1] With chronic kidney disease, high cardiovascular morbidity and mortality rates are prevalent Approximately 25–30 % of all diabetics eventually develop kidney damage that mostly

is of progressive nature [2 3] Diabetic nephropathy is the major cause of end-stage renal disease in Western societies; nevertheless, aetiology and pathogenesis of the observed kidney damage are incompletely understood Differential diagnosis for chronic kidney diseases in diabetics is ample, given that arterial hypertension is highly prevalent and may

Abstract

Background and objective In diabetics genetic

predis-position, poor glycemic control and arterial

hyperten-sion contribute to nephropathy development in patients

affected by diabetes mellitus We set up the hypothesis

that diabetic nephropathy and incisional hernia formation

may have in common alterations of collagen composition

and tested whether the occurrence of diabetic nephropathy

coincides with wound healing disturbance (incisional

her-niation) or connective tissue diseases (inguinal herniation,

umbilical herniation, aortic aneurysm, varicose veins, disc

herniation)

Design A questionnaire on surgical procedures, wound

healing and connective tissue disorders was performed

with 731 diabetics Furthermore, test results for kidney

function and damage (creatinine clearance, proteinuria)

and blood sugar control (HbA1c) were recorded

Correla-tions between aforementioned connective tissue diseases

and “advanced” diabetic nephropathy were calculated

“Advanced” diabetic nephropathy was assumed in patients

with macroproteinuria, CKD stage 5 and/or end-stage renal

disease All diabetics with CKD stages 1 and 2 without

pro-teinuria were included in the “control” group A subgroup

analysis on incisional hernia formation coinciding with

diabetic nephropathy was performed in patients with

previ-ously performed abdominal surgery

Results In patients with advanced nephropathy, some

dis-eases with connective tissue alterations, such as inguinal

* Paula Loewe

paula.holstiege@med.ovgu.de

1 Department of Nephrology and Hypertension, Diabetes

and Endocrinology, Otto-von-Guericke-University

Magdeburg, Leipziger Str 44, 39120 Magdeburg, Germany

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contribute to the pathogenesis of kidney alterations and/or

itself cause nephrosclerosis Furthermore, it is well known

that about 30–40 % of diabetics suffer from other kidney

diseases than diabetic nephropathy [4 5] Without retrieval

of kidney tissue, such a differentiation is not possible with

certainty; furthermore, the range of proteinuria may only

provide a hint at the extent of glomerular damage In recent

years, several studies report on a subgroup of patients with

progressive diabetic nephropathy that lack significant

pro-teinuria at any stage of disease [6 7] Notwithstanding, the

hypothesis to be tested in this project is a deranged

under-lying collagen metabolism due to genetic traits favouring

progressive kidney disease These tests are similarly

appli-cable in cohorts encompassing nephrosclerosis or other

(immunological) kidney diseases Main predictors of

pro-gression to end-stage renal disease (ESRD) include genetic

predisposition [8 9], an insufficiently controlled glucose

metabolism [10], arterial hypertension, dyslipidemia and

nicotine consumption [11, 12]

In order to prevent and treat incipient and overt diabetic

nephropathy, understanding of the underlying pathogenesis

is essential The focus of different research groups has been

a dysregulated inflammatory response, matrix composition

or regeneration of connective tissue [13, 14] Our research

group and others found alterations of the collagen

com-position of kidneys in patients with diabetic nephropathy

similar to those of the scar tissue of patients with incisional

herniation [15–17], where the ratio of collagen type I and

collagen type III is balanced in favour of the less stable

collagen type III fibrils These observations led us to the

question if alterations in wound healing contribute to the

pathogenesis of diabetic nephropathy and/or if impaired

wound healing is a risk factor for diabetic nephropathy

development

Chronic connective tissue diseases, such as varicose

veins, aortic aneurysms, inguinal herniation, umbilical

her-niation and disc herher-niation, exhibit an altered extracellular

matrix [18] Some of them are associated with wound

heal-ing disorders [19] Patients with abdominal aortic

aneu-rysms that undergo reconstructive surgery more often suffer

from incisional herniation than patients undergoing other

operations [19] Bode et al [20] detected a raised turnover

of collagen type III in the aortic vessel wall of abdominal

aneurysms by immunohistochemistry Fachinelli et al [15]

found a decrease in total collagen and collagen type I in

the linea alba of patients with ventral herniation Raffeto

et al compared patients with abdominal aortic aneurysms

to patients with aortoiliac occlusive disease undergoing

sur-gery Patients with abdominal aortic aneurysms more

com-monly had abdominal wall and inguinal herniations as well

as a higher risk of developing an incisional herniation [21]

Given that most of the diabetics suffering from

nephropathy also develop arterial hypertension, it may be

envisioned that repetitive elevated intraglomerular pressure

is translated into glomerular injury that has to be confined Reparative processes within the glomerular architecture have been described in diabetics [1] Classifications on dia-betic nephropathy, such as the one by Taervert et al [22], describe glomerular fibrosis and mesangial cell prolifera-tion features resembling a wound healing process

Materials and methods

Patient enrolment and data collection took place at a ter-tiary medical centre (University Hospital Aachen), and out-patient private practices specialized in Diabetes and Neph-rology The ethical committee of the University Hospital RWTH Aachen approved the study (EK Nr 68/09) Upon informed written consent, standardized interviews were performed with patients that address wound healing (inci-sional herniation) and connective tissue (inguinal hernia-tion, umbilical herniahernia-tion, aortic aneurysm, varicose veins and disc herniation) disorders Laboratory testing included serum creatinine, HbA1c and proteinuria (daily protein excretion rate in 24-h urine samples) Information on bio-metric data (gender, age, BMI) and known risk factors for the development of diabetic nephropathy (diabetes mellitus duration, smoking habits, arterial hypertension and dys-lipidemia/usage of lipid-lowering drugs) and risk factors for incisional hernia development [angiotensin-converting enzyme (ACE)-inhibitor/angiotensin receptor (AT)1-antag-onist medication, non-steroidal anti-inflammatory drug (NSAID) intake, history of glucocorticoid intake and inci-dence as well as number of previous abdominal surgeries] were collected Estimated glomerular filtration rate (eGFR) was calculated by the Modification of Diet in Renal Dis-ease (MDRD) formula

Amongst the 731 patients enrolled in the study those with macroproteinuria (>300 mg/day), impaired eGFR < 15 ml/min/1.73 m2 or dialysis dependency was regarded as “advanced” diabetic nephropathy For statisti-cal analyses and as comparator, a control group was defined which consisted of diabetics with an eGFR > 60 ml/ min/1.73 m2 and lack of proteinuria (<30 mg/day) The diabetics with eGFR > 15 ml/min/1.73 m2 and <60 ml/ min/1.73 m2 or those with eGFR > 60 ml/min/1.73 m2

and microalbuminuria were excluded for the initial main statistical analyses and only used as comparator group in

a subanalysis for disc herniation Kidney biopsy for diag-nosis of diabetic nephropathy was not enforced, given that this is not commonly accepted due to potential com-plications (bleeding, infection) and the lack of therapeutic consequences

A subgroup analysis regarding the correlation of inci-sional hernia formation and advanced diabetic nephropathy

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was performed in patients that previously underwent

abdominal surgery (n = 381).

Since patients on dialysis are bound to rest for

pro-longed periods during renal replacement therapy more

fre-quently, the tested relationship between advanced diabetic

nephropathy and herniated discs may be confounded Thus,

for comparison a group consisting of diabetics with an

eGFR between 15 and 60 ml/min/1.73 m2 or

microprotein-uria (30–300 mg per day) was defined in order to compare

patients without nephropathy and moderate,

non-dialysis-dependent nephropathy with regard to disc herniation

Statistics

Metrical variables were assessed for normal distribution

using the Kolmogorov–Smirnov test Depending on the data

distribution, t tests or Mann–Whitney U tests were used for

group comparisons Nominal data were examined using

chi-square test Since the development of diabetic

nephropa-thy is subject to several confounding factors, multivariate

analyses were performed Therefore, binary logistic

regres-sion was executed for independence of risk factors Known

risk factors or possible new modifiers were considered if

reaching a p value < 0.1 in univariate analysis

Addition-ally, known risk factors for the genesis of incisional hernias

were included A p value <0.05 was regarded as statistically

significant The statistical analysis was accomplished using

SPSS (version 15.0, IBM SPSS Inc., Somer, NY, USA)

Results

Following informed written consent, 731 diabetics were

enrolled in the study: 410 (56.1 %) of these were male

and 321 (43.9 %) female Kidney function was calculated

above an estimated glomerular filtration rate of 60 ml/

min/1.73 m2 in 353 patients, whereas 212 patients suffered

from moderate (stages III/IV or microalbuminuria) and

141 from advanced nephropathy defined by dependency on

dialysis, CKD stage V or macroalbuminuria

Diabetics with “advanced” nephropathy were older, had

a lower HbA1c (6.5 vs 7.3 %), were diagnosed for a longer

time period with diabetes mellitus, suffered more often from arterial hypertension, smoked less frequently and took lipid-lowering drugs more often (Table 1) Some connec-tive tissue diseases (inguinal herniation, aortic aneurysm and varicose veins) were similarly often diagnosed in the control group as well as in those patients with “advanced” nephropathy (Table 2) Diabetics with preserved kid-ney function (eGFR > 60 ml/min/1.73 m2) reported about umbilical herniation more often than patients with

“advanced” nephropathy (Table 2) Disc herniation is sig-nificantly less often diagnosed in diabetics with “advanced” nephropathy (Fig 1; Table 2) Comparing diabetics with normal kidney function to those with moderately impaired kidney function (eGFR 15–60 ml/min/1.73 m2 or micro-proteinuria) revealed that disc herniation is significantly

less prevalent in the latter group (8 vs 16.1 %, p = 0.008;

Figs 1 2)

Abdominal surgery had previously been performed in

381 patients Appendectomy (n = 158) was by far the most common operation, followed by hysterectomy (n = 58), cholecystectomy (n = 57) and inguinal hernia opera-tion (n = 56) Gynaecological operaopera-tions other than

hys-terectomies were recorded in 74 cases Eighteen patients had received kidney or kidney and pancreas transplants Subgroup analysis of these patients demonstrated inci-sional hernia formation was significantly more frequent

Table 1 Factors influencing

the development of “advanced”

diabetic nephropathy

Normal kidney function

n = 353

“Advanced” kidney disease

n = 141

p value

Diabetes duration ≥ 10 [n (%)] 184 (52.4 %) 103 (75.2 %) <0.001

Table 2 Outcome regarding connective tissue disorders of diabetics

with preserved kidney function and “advanced” kidney disease

Preserved kidney function

n = 353

“Advanced”

kidney disease

n = 141

p value

Aortic aneurysm

[n (%)]

Varices [n (%)] 80 (22.7 %) 26 (18.4 %) 0.3 Inguinal hernia

[n (%)]

Umbilical herniation

[n (%)]

Disc herniation

[n (%)]

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when “advanced” nephropathy was concomitantly present (16.0 %), whereas only 5.7 % of patients suffered from incisional herniation without “advanced” nephropathy The potential risk factors for the development of inci-sional herniation considered in the statistical analysis are shown in Table 3 A significant association in the univariate analysis could only be found between kidney function and incisional hernia formation as well as a history of glucocor-ticoid treatment and incisional hernia formation

Variables considered in multivariate analysis were age, duration of diabetes mellitus since diagnosis, nicotine consumption, arterial hypertension, usage of lipid-low-ering drugs, incisional hernia formation and therapy with glucocorticoids

By means of binary logistic regression, we could iden-tify incisional herniation as an independent risk factor (OR

3.5, p = 0.022) for the development of “advanced” diabetic

nephropathy Other independent risk factors were age (OR

2.6, p = 0.04), diabetes mellitus duration since diagnosis (OR 2.5, p = 0.019) and arterial hypertension (OR 1.1,

p < 0.001, Table 4)

Discussion

The pathophysiology of diabetic nephropathy is still not resolved, although enormous effort has been made to elu-cidate common underlying modes of damage [23, 24] The coincidence of arterial hypertension may aggravate kidney disease The mainstay of histological alterations consists of thickened basement membranes; furthermore, a pathologi-cal classification has been described that is based on glo-merular changes [22]

The initial hypothesis of our study was that diabetic nephropathy and incisional hernia development may have similar underlying pathophysiology, that is, a tissue regen-eration disorder with altered collagen metabolism and com-position For recurring incisional hernia formation excess collagen type III over collagen type I expression has been described [25]; similarly, alterations of collagen synthesis have been described for kidney fibroblasts grown under hyperglycaemic conditions [26, 27] However, based on the data on hand, it is not possible to give a statement con-cerning the causality for disease onset and perpetuation The tensile strength of type III collagen differs consider-ably from type I collagen, which is the main constituent of mature scar tissue Wound healing may be skewed by such dysregulations [28]; furthermore, loss of organ function may be incited by recurring injuries and misguided wound

83.1%

92%

94.3%

16.9%

8%

5.7%

0

50

100

150

200

250

300

350

CKD stage I/II, no

albuminuria CKD stage III/IV ormicroalbuminuria CKD stage V and/ormacroalbuminuria

n

no disc herniation disc herniation

Fig 1 Frequency of disc herniation in dependency of kidney

func-tion The share of diabetics with disc herniation was higher in

patients with preserved kidney function and lower in patients with

moderately or markedly “advanced” kidney disease

94.3%

84.0%

0

20

40

60

80

100

120

140

160

180

200

CKD stage I/II no

albuminuria CKD stage V and/or macroalbuminuria

n

no incisional herniation incisional herniation

Fig 2 Frequency of incisional herniation in dependency of kidney

function The percentage of patients with incisional herniation in the

group with preserved kidney function demonstrates a lower share of

5.7 % when compared with those with “advanced” kidney disease

(16.0 %)

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healing One may simplify the underlying

pathomecha-nism as wounding with ensuing wound repair that results in

immature scarring and excessive collagenous tissue

deposi-tion in the kidneys

Our statistical analysis revealed an association between

“advanced” diabetic nephropathy and incisional hernia

development within the subgroup of patients that had a

pre-ceding abdominal surgery Diabetics with a positive past

medical history for incisional abdominal herniation had an

increased relative risk of 3.5-fold for the development of

“advanced” diabetic nephropathy Several endogenous

fac-tors (age, gender, obesity, anaemia and diabetes mellitus) as

well as exogenous factors (glucocorticoids, ACE-inhibitors,

NSAID and smoking) promote incisional hernia

forma-tion [29] Our study did not identify a significant difference

within the gender distribution and the body mass index

(BMI) was not higher in patients with incisional abdominal

herniation The number of preceding abdominal operations

was about the same in both groups, the ones with and

with-out incisional hernia formation

Our study shows a borderline significant correlation

between ACE-inhibitor or AT-1 receptor antagonist intake

and the occurrence of incisional herniation (p = 0.07)

Uni-variate analysis showed an association between the usage

of glucocorticoids and a history of incisional herniation

Both did not reach levels of significance in the multivariate analysis

By multivariate analysis, we identified incisional her-niation as independent risk factor for “advanced” diabetic nephropathy development In addition, diabetic nephropa-thy was independently associated with age, duration of dia-betes mellitus and arterial hypertension

Patients with end-stage kidney disease have complex mechanisms involved in poor wound healing [30, 31]

We therefore performed another statistical analysis with patients suffering from CKD and macroproteinuria, exclud-ing those with end-stage kidney disease A significant posi-tive association between advanced diabetic nephropathy

and incisional herniation rates (p = 0.002) was still found,

whereas with other connective tissue alterations, no corre-lations were present

Thus, our results support the assumption that both dis-eases underlie similar pathomechanisms leading to an impaired regeneration of connective tissue

The gene expression profile of fibroblasts outgrown from the skin of patients with inguinal hernias also exhib-ited a reduced ratio of type I/III collagen [18] Neverthe-less, our data do not provide a positive correlation between inguinal herniation and “advanced” diabetic nephropathy development

Umbilical herniation was less frequent in diabetics with advanced nephropathy (3.0 %) compared to the cohort of patients with preserved kidney function (8.2 %) Fach-inelli et al showed a reduction in total collagen as well as

a reduced collagen type I in the linea alba of patients with ventral herniation (umbilical, epigastric and incisional her-niation) compared to healthy controls Differences in the amount of collagen type III were not described [15] Varia-tions in the composition of the connective tissue in the dif-ferent types of herniation were not considered

Table 3 Factors influencing

the development of incisional

herniation

a Preserved kidney function

b Advanced kidney disease

No incisional herniation n = 224 Incisional hernia n = 19 p value

Number of previous operations [median]

Kidney function

Gender

Table 4 Binary logistic regression with diabetic nephropathy risk

factors

p univariate OR (CI 95 %) p multivariate Incisional herniation 0.016 3.5 (1.2–10.3) 0.022

Diabetes duration <0.001 2.5 (1.2–5.3) 0.02

Hypertension <0.001 1.1 (1.0–1.1) <0.001

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Furthermore, disc herniation was less frequent in

diabet-ics with “advanced” nephropathy One possible

explana-tion is that patients undergoing renal replacement therapy

have to rest in supine position for several hours during each

dialysis session and may thus be protected from spine

pres-sure expopres-sure However, a similar negative correlation was

also found for patients that had not yet progressed to

end-stage renal disease and suffered from moderately impaired

kidney function (defined as eGFR 15–60 ml/min/1.73 m2,

microproteinuria) when compared to diabetics with normal

kidney function The pathomechanism of diabetic

nephrop-athy and disc herniation seem to be opposing; that is,

patients with “advanced” nephropathy were protected from

disc herniation From experimental and genetic studies, it

is known that matrix metalloproteinase (MMP) activities

are lowered in diabetic nephropathy [32] The gene

expres-sion of gelatinase B is reduced when a poorly controlled

diabetic metabolism prevails and glomerulosclerosis

coex-ists [33] A dinucleotid repeat polymorphism within the

matrix metalloproteinase-9 (MMP-9 or gelatinase B) gene

was found, the presence of which was associated with

dia-betic nephropathy [34] Conversely, increased activities

of gelatinase and stromelysin activities were detected in

humans with lumbar and cervical disc herniation [35, 36]

Dogs with acute disc herniation exhibited elevated

gelati-nase B enzymatic activity in the cerebrospinal fluid [37]

Park et al [38] showed that the concentration of the

tis-sue inhibitor of matrix metalloproteinase 2 (TIMP-2) was

lower in the ligamentum flavum of patients with disc

herni-ation when compared to the ligamentum flavum of patients

with spinal stenosis

Our study has several limitations The data collection

has been standardized; however, it has been performed in

a retrospective manner by questionnaires The follow-up

and observation period since diagnosis of diabetes mellitus

differed considerably for the recruited patients and ideally

should be at least 15 years, a time period when the

inci-dence of diabetic nephropathy development peaks

Fur-thermore, the “classification” as diabetic nephropathy may

not be justified, given that a considerable share of patients

suffers from coinciding other kidney pathologies that may

not be excluded without performance of kidney biopsies

The frequencies of normoalbuminuria observed in patients

with low GFR have been reported to be 22–24 % in type

1 diabetes mellitus and 32–71 % in type 2 diabetes

melli-tus [39] Thus, our classification with macroalbuminuria as

indicator of “advanced” diabetic nephropathy may not be

justified in all patients; however, it seems the best available

indicator of kidney damage besides impaired glomerular

filtration rate The study may be subject to various forms

of bias: 1 Reporting bias As the primary end-point

(her-nia, etc.) is based on a standardized interview, some cases

may be overlooked, others falsely included 2 Selection

bias Only patients admitted to the surgery ward of a ter-tiary medical centre were approached and asked for partici-pation in the interview Most of the patients were enrolled; however, some denied participation without providing rea-sons 3 Information bias It is not clear whether the sur-gical procedures (e.g appendectomy) were performed at a time when the patient fulfilled the criteria for (advanced) diabetic nephropathy Hence, some patients may have been misclassified 4 Confounding bias The effects of advanced diabetic nephropathy may be confounded by different BMI/ medication/diet/occupation However, BMI for the patients with and without incisional herniation was similar (see Table 3) The healthy worker effect Patients with diabetes mellitus and stage I/II nephropathy are by far more likely to have different occupations than patients with stage V All these effects may be operative in the study and may confound our results The standardized interview has been chosen to minimize differences in questionnaires, and only

a single person performed the questionnaire to exclude dif-ferences in interviewing between groups

Conclusion

Our study strongly underscores that in diabetics connec-tive tissue composition and related diseases share under-lying pathophysiology that may affect integrity of kidney architecture and function The occurrence of incisional herniation positively correlates with “advanced” kidney disease development, whereas umbilical herniation and disc herniation indicate “protection” from perpetuated kid-ney damage

Acknowledgments We are thankful for the collaboration and

sup-port with patient recruitment by Dr Heddeus, Dr Weidemann and Dr Böhm.

Funding The study was supported by SFB/TR57 (Project TP4),

SFB854(TP01) and Me1365/9-1 to PRM.

Compliance with ethical standards Conflict of interest The results presented in this paper have not been

published previously in whole or part, except in abstract format.

Ethical approval The study protocol was approved by the

ethical-committee of the RWTH University Aachen.

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